Inhibins and activins are members of the transforming growth factor beta (TGF?) family of cytokines. Inhibins are heterodimers consisting of a common ?-subunit linked to either a ?A subunit ( ?-?A, forming inhibin A) or a ?B subunit ( ?-?B, forming inhibin B). Activins share the ?-subunit with the inhibins and may be homo or heterodimers of ?-subunits forming activin A (?A-?A), activin AB (?A-?B) or activin B (?B-?B). The expression of the ?-subunit, and therefore of inhibins appears to be more restricted than that of the ?-subunit, and therefore of activins. Inhibins and activins play a role in the regulation of pituitary follicle stimulating hormone (FSH) secretion.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
AMY82
Concentration:
Greater than or equal to 214 mg/L
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
Robertson D et al. Endocrine-Related Cancer. 2004; 11:3549
References 2:
Bernard J et al. Recent Progress in Hormone Research. 2001; 56:417450
Tumor cells of epithelial, lymphoid, glial and mesenchymal origin are reported to be negative. This clone is well described in the literature. It is indicated to label an intracytoplasmic antigen in the majority of melanomas and other tumors demonstrating melanoma/melanocytic differentiation.
The CD123 antigen is also known as the alpha subunit of the human interleukin-3 receptor. It is a type I transmembrane glycoprotein and is a member of the cytokine receptor superfamily. CD123 forms a heterodimer with CD131 (the beta subunit of the interleukin-3 receptor) to form the interleukin-3 receptor, where the cytokine specificity is provided by the alpha subunit and the signal transduction function is provided by the beta subunit. The interleukin-3 receptor is reported to be expressed on monocytes, neutrophils, basophils, eosinophils, megakaryocytes, erythroid precursors, mast cells, macrophages and a subpopulation of B cells, where it mediates proliferation and differentiation of these cells. Outside the hematopoietic system CD123 is reported to be expressed in Leydig cells of the testis, some endothelial cells, and cells of the placenta and brain.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
BR4MS
Concentration:
Greater than or equal to 90 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
GarnacheOttou F et al. British Journal of Haematology. 2007; 136:539548
References 2:
Moretti S et al. J.of Biol.Regulators and Homeostatic Agents. 2001; 15:98100
CD99 is a 32 kDa transmembrane glycoprotein, encoded by the MIC2 gene, which is located in the pseudoautosomal region of the human X and Y chromosomes. Recently, the MIC2 gene has been shown to encode two distinct proteins which are produced by alternative splicing of the CD99 gene transcript and are identified as bands of 30 and 32 kDa (p30/32).
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
PCB1
Concentration:
Greater than or equal to 9,9 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Bühnemann C et al. PLoS ONE. 2014; 9(9): e107105.
References 2:
Bahrami A et al. Archives of Pathology and Laboratory Medicine. 2008; 132:326-348
Calcitonin (CT) is a 32 amino acid peptide synthesized by the parafollicular C cells of the thyroid. It acts through its receptors to inhibit osteoclast mediated bone resorption, decrease calcium resorption by the kidney and decrease calcium absorption by the intestines. The action of calcitonin is therefore to cause a reduction in serum calcium, an effect opposite to that of parathyroid hormone. The calcitonin gene transcript also encodes the calcitonin gene-related peptide (CGRP), which is thought to be a potent vasodilator. The tissue specificity of the transcript produced depends on alternative splicing of the CT/CGRP gene transcript. In the parafollicular cells of the thyroid 95% of the CT/CGRP is processed and translated to produce CT, however, in neuronal cells 99% of the CT/CGRP RNA is translated into CGRP. The C cells of the thyroid give rise to an endocrine tumor, medullary thyroid carcinoma (MTC), which occurs in a sporadic (75% of cases) and hereditary form (25% of cases). Familial MTC is associated with C cell hyperplasia (CCH), whereas sporadic MTC is thought not to be. However, in the general population CCH is present in 20-30% of thyroid glands, either with normal histology, thyroiditis or follicular tumors.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
CL1948
Concentration:
Greater than or equal to 29 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Leboulleux S et al.Clinical Endocrinology. 2004; 61:299310
References 2:
Hirsch P et al. Journal of Musculoskeletal & Neuronal Interactions. 2001; 1(4):299305
References 3:
Pondel M. International Journal of Experimental Pathology. 2000; 81:405422
The MUM-1 (multiple myeloma oncogene 1) gene was originally identified because of its involvement in the t(6:14) translocation observed in multiple myeloma, which causes the juxtaposition of the MUM-1 gene to the Ig heavy chain locus. MUM-1 is expressed in late plasma cell directed stages of B cell differentiation and in activated T cells, suggesting that MUM-1 may serve as a marker for lympho-hemopoietic neoplasms derived from these cells. The morphologic spectrum of MUM-1 expressing cells has been found to range from that of a centrocyte to that of a plasmablast/plasma cell. Consequently the histogenic value of MUM-1 may be to provide a marker to aid in the identification of the transition from BCL-6 positive (germinal center B cells) to CD138 positive (immunoblasts and plasma cells).
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
EAU32
Concentration:
Greater than or equal to 263 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Bergsagel P and Kuehl W.Oncogene. 2001: 20(40);5611-5622
Alpha-methylacyl-CoA racemase (AMACR), also known as p504s, is a mitochondrial and peroxisomal enzyme that is involved in bile acid biosynthesis and beta-oxidation of branched-chain fatty acids. AMACR is essential in lipid metabolism, and is expressed in normal liver (hepatocytes), kidney (tubular epithelial cells) and gall bladder (epithelial cells). Expression has also been found in lung (bronchial epithelial cells) and colon (colonic surface epithelium). Expression is granular and cytoplasmic. AMACR expression can also be found in hepatocellular carcinoma and kidney carcinoma. Past studies have also shown that AMACR is expressed in various colon carcinomas (well, moderately and poorly differentiated) and over expressed in prostate carcinoma.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
EPMU1
Concentration:
Greater than or equal to 84 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Lloyd M et al.FEBS Journal. 2008: 275;10891102
References 2:
Rubin M et al. J. of the Am. Med.Assoc. 2002: 287(13);16621670
The basic structure of an immunoglobulin molecule consists of two identical heavy chains, either gamma, alpha, delta, or epsilon and two identical light chains, either kappa or lambda. Any heavy chain can associate with either light chain but on any immunoglobulin molecule both light chains are of the same type. The ratio of kappa and lambda light chains varies between Ig classes and subclasses. In a polyclonal population the ratio of kappa to lambda bearing B cells is approximately 2:1, with individual B cells thought to express kappa or lambda light chains, never both. The majority of kappa and lambda chains are bound to heavy chain immunoglobulin, however in normal individuals low levels of free light chain are present in serum. The occurrence of a mixture of kappa and lambda chain expressing cells suggests a polyclonal population and a reactive or non-neoplastic proliferation of B cells.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
SHL53
Concentration:
Greater than or equal to 554 mg/L
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
Gertz M et al. Kidney International. 2002; 61(1):19
References 2:
Ramsland P and Farrugia W. Journal of Molecular Recognition. 2002; 15:248259
Immunoglobulins are polypeptides and comprise five major classes; immunoglobulin G (IgG), IgA, IgM, IgD and IgE. Each immunoglobulin consists of two identical heavy (H) chains and two identical light (L) chains.These are also subdivided into sub classes eg IgG1. There are two classes of light chain; kappa and lambda. The ratio of kappa chains and lambda chains varies between Ig classes and sub classes, but is also species specific. In humans, approximately 60 percent of light chains are kappa. However, in any particular immunoglobulin molecule the light chain will be either kappa or lambda. B cells contain either kappa or lambda mRNA.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
CH15
Concentration:
Greater than or equal to 125 mg/L
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
Ramsland P and Farrugia W. Journal of Molecular Recognition. 2002; 15:248259
Folate is a basic component of cell metabolism and DNA synthesis and repair. It is involved in essential one-carbon transfer reactions and is a vitamin required by both normal and tumor cells. Folate entry into cells is facilitated via two different systems: the reduced folate carrier, which utilizes a bidirectional anion-exchange mechanism, and the folate receptor system. Folate receptor alpha is a membrane-bound member of the folate receptor family, facilitating folate transport via a mechanism termed potocytosis where the receptor is internalized and then recycled back to the cell membrane. Folate receptor alpha expression is reported to be highly restricted in normal tissues and only selectively overexpressed in a limited number of epithelial malignancies.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
BN3.2
Concentration:
Greater than or equal to 67 mg/L
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
Smith AE et al. Hybridoma. 2007; 26(5):281288
References 2:
Kelemen L. International Journal of Cancer. 2006; 119:243250
IgM, together with IgD, is the major immunoglobulin expressed on the surface of B cells and normally constitutes about 10 per cent of serum immunoglobulin. IgM antibody is prominent in early immune responses to most antigens and predominates in certain antibody responses such as natural blood group antibodies.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
8H6
Concentration:
Greater than or equal to 41 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Sangaletti S et al. Oncoimmunology. 2014; 3:e28989. doi: 10.4161/onci.28989
References 2:
Carsetti R et al. Immunological Reviews 2004; 197: 179-191
References 3:
Schaffer A et al. Nature Reviews: immunology. 2002; 2: 1-13
Mouse anti-Delta chain of human Immunoglobulin D, clone DRN1C (monoclonal)
Antibody Type:
Monoclonal
Host Animal:
Mouse
Species Reactivity:
human
Immunogen:
Prokaryotic recombinant protein corresponding to 222 amino acids of the N terminus of the delta heavy chain constant region of the human immunoglobulin D molecule.
IgD, together with IgM, are the major immunoglobulins expressed on the surface of B cells where it seems they may operate as mutually interacting antigen receptors for the control of lymphocyte activation and suppression. The greater susceptibility of IgD to proteolysis in combination with antigen could well be implicated in such a function. The use of PBS-based diluents may result in increased background staining. Clone DRN1C was developed to produce reduced background staining that is associated with polyclonal antibodies on paraffin sections.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
DRN1C
Concentration:
Greater than or equal to 133 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Geisberger R et al. Immunology. 2006; 118:429-437
References 2:
Preudhomme J et al. Molecular Immunology. 2000; 37:871-887
References 3:
Vladutiu A. Clinical and diagnostic laboratory immunology. 2000; 7(2):131-140
IgA is a member of the antibody class of the immunoglobulin superfamily. There are several classes and subclasses (isotypes) of antibody, the antibody isotype being defined by the immunoglobulin heavy chain present in the molecule. The basic structure of an immunoglobulin molecule consists of two identical heavy chains (gamma , mu, alpha , delta , epsilon) and two identical light chains, either kappa or lambda. IgA contains the alpha -chain and may be present in a serum or secretory form. In serum, 90% of IgA is monomeric, while in its secretory form it is the main immunoglobulin found in secretions including tears, saliva, intestinal and bronchial mucous, sweat, colostrum, and secretions from the prostate and respiratory epithelia, where it has the job of defending exposed external surfaces of the body against attack from micro organisms. Secretory IgA is synthesized locally by plasma cells and dimerized intracellularly with a cysteine-rich J-chain. Clone N1CLA was developed to produce reduced background staining that is associated with polyclonal antibodies on paraffin sections.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
N1CLA
Concentration:
Greater than or equal to 46 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Merluzzi S et al. Blood Journal. 2010; 115(14):2810-2817
References 2:
Fagarasan S and Honjo T. Current opinion in Immunology. 2004; 16(3):277-283
References 3:
Pilette C et al. European Respiratory Journal. 2001; 18:571-588
Polycomb-group proteins (PcG) such as EZH2 (Enhancer of Zeste Homolog 2 (Drosophila)) form multimeric gene repressing complexes involved in axial patterning, hematopoiesis and cell cycle regulation. PcG proteins ensure correct embryonic development by expressing homeobox genes as well as contributing to the regulation of lymphopoiesis.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
6A10
Concentration:
Greater than or equal to 20 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Van Kemenade FJ et al. Blood, 97(12): 38963901 (2001)
References 2:
Raaphorst Fm et al.American Journal of Pathology, 157(3): 709715 (2000)
References 3:
Kattan M. Journal of the National Cancer Institute. 95(9): 634635 (2003)
DOG-1, a 986 amino acid protein of unknown function, is expressed predominantly on the plasma membrane of gastrointestinal stromal tumors (GISTs) and is rarely expressed in other soft tissue tumors, which, due to appearance, can be confused with GISTs. Reactivity for DOG-1 has been suggested to aid in the identification of GISTs, including Platelet-Derived Growth Factor Receptor Alpha mutants that fail to express KIT antigen. The use of PBS-based diluents may result in increased background staining.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
K9
Concentration:
Greater than or equal to 12 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Novelli M et al. Histopathology. 2010; 57, 259-270
References 2:
Miettinen M et al. American Journal of Surgical Pathology, 2009; 33, 1401-1408
The CD30 antigen is a single chain glycoprotein with a molecular weight of 120 kD. CD30 antigen is known to act as a receptor for a cytokine ligand, CD30L, and may also play a role in the regulation of cellular growth and transformation. CD30 antigen is reported to be expressed on the surface of multinucleated Reed Sternberg cells, mononuclear Hodgkin's cells and in the majority of anaplastic large cell lymphomas. The CD30 antigen is expressed in non-Hodgkin's lymphoma and virally transformed cells, for example, EBV-transformed B cells.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
JCM182
Concentration:
Greater than or equal to 72 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Kennedy M et al. Immunology. 2006; 118: 143-152
References 2:
Chiarle R et al. Clinical Immunology. 1999; 90(2):157-164
CD19 is a member of the immunoglobulin superfamily and has two Ig like domains. It is a single chain glycoprotein present on the surface of B lymphocytes and follicular dendritic cells of the hematopoietic system. CD19 is a crucial regulator in B cell development, activation and differentiation. On B cells, CD19 associates with CD21, CD81 and CD225 (Leu-13) forming a signal transduction complex. CD19 is expressed from the earliest recognizable B cell lineage stage, through development to B cell differentiation but is lost on maturation to plasma cells.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
BT51E
Concentration:
Greater than or equal to 35 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Fujimoto M and Sato S. Journal of Dermatological Science. 2007; 46:1-9
References 2:
Otero D et al. The Journal of Immunology. 2003; 170: 73-83
References 3:
Fujimoto M et al. Seminars in Immunology. 1998; 10:267-277
Carbonic anhydrase (CA) is an enzyme that assists rapid interconversion of carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. Originally named MN/G250, carbonic anhydrase IX (CAIX) is a cell surface transmembrane protein, which is predominantly found in the gastrointestinal tract and gallbladder. The glandular regions of normal colon are reported to be negative, but in the case of adenocarcinoma, the glands are positive. CAIX is also reported to be expressed in common epithelial tumors such as carcinomas of the esophagus, lung, colon, kidney, cervix and non-small cell lung carcinoma.In breast carcinomas, CAIX expression has been reported to be associated with malignant tissue. Expression of CAIX is reported to be absent in normal kidney, chromophobe carcinomas or oncocytomas; however, it is specifically expressed in clear cell renal carcinomas.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
TH22
Concentration:
Greater than or equal to 21 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Swietach P et al. Cancer and Metastasis Reviews. 2007; 26:299310
References 2:
Potter C and Harris A. Cell Cycle. 2004; 3(2):164167
Akt-1, also referred to as protein kinase B (PKB) or Rac alpha is a member of the Akt serin/threonine protein kinase family. It plays an important role in many biological responses including metabolism, cell survival and growth by phosphorylation and inactivating several targets including GSK 3 beta, caspase 9, BAD and the forkhead transcription factor. Akt-Phos is not recommended for use with PBS, since the use of PBS-based wash buffers and possibly PBS-based antibody diluents gives increased background staining and decreased staining intensity.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
LP18
Concentration:
Greater than or equal to 300 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Brazil D et al. Trends in Biochemical Sciences. 2004; 29(5):233242
References 2:
Nicholson K et al. Cellular Signalling. 2002; 14:381395
References 3:
Lawlor M et al. Journal of Cell Science. 2001; 114:29032910
ZAP-70 is a member of the syk family of proteins. It is expressed on T cells and NK cells and is required for the T cell receptor activation that triggers an immune response. CLL B cells that express the non-mutated immunoglobulin VH genes express levels of ZAP-70 protein that are comparable to those found in the blood T cells of healthy adults. Leukemic cells that express mutated Ig VH genes generally do not express detectable levels of ZAP-70 protein and this is correlated with the high level expression of CD38.
Antibody Isotype:
IgG2b, kappa
Monosan Range:
MONXtra
Clone:
L453R
Concentration:
Greater than or equal to 449 mg/L
Storage buffer:
PBS with sodium azide
Storage:
2-8°C
References 1:
Orchard J et al. Leukaemia and Lymphoma 2005; 46(12):16891698
References 2:
Wang J et al. Applied Immunohistochemistry and Molecular Morphology 2005; 13(4):323332
References 3:
Mustelin T and Tasken K.Biochemical Journal 2003; 371:1527
References 4:
Van Oers N and Weiss A. Seminars in Immunology 1995; 7:227236
Eukaryotic cells contain a number of types of cytoplasmic filamentous proteins, microtubule, microfilaments and intermediate-sized filaments (IF). Vimentin, a 57 kD protein that is an intermediate filament is reported to be expressed in most cells of mesenchymal origin, including fibroblasts, endothelial cells, smooth muscle, melanocytes as well as T and B lymphocytes.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
SRL33
Concentration:
Greater than or equal to 60 mg/L
Storage buffer:
Tissue culture supernatant with 15 mM sodium azide
Storage:
2-8°C
References 1:
Runembert I et al. Journal of Cell Science 115: 71324 (2002)
Thyroid peroxidase gene expression is under the regulation of thyroid stimulating hormone. In normal thyroid, expression of thyroid peroxidase (TPO) described immunohistochemically is reported to produce a diffuse, fine, granular cytoplasmic stain in all follicular cells. Some studies have shown qualitative, as well as quantitative differences in thyroid peroxidase expression in thyroid cancer compared to normal tissue.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
AC25
Concentration:
Greater than or equal to 10.8 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Kimura S et al. Proceedings of the National Academy of Science USA 84: 5555-5559 (1987)
References 2:
De Micco C et al. Cancer 67(12): 30363041 (1991)
References 3:
Czarnocka B et al. Breast Journal Cancer 85(6): 875880 (2001)
References 4:
Tanaka T et al. Journal of Pathology 179: 8994 (1996)
Oct-3/4 is a member of the POU homeodomain family of transcription factors, which is expressed by embryonic stem cells and germ cells. A critical amount of Oct-3/4 is required to maintain stem cell self replication. Down regulation of Oct-3/4 levels are associated with loss of pluripotency. Oct-3/4 has been proposed as a useful marker for germ cell tumors which exhibit features of pluripotentiality, including seminoma/dysgerminoma/germinoma and embryonal carcinoma, and establishing a germ cell origin for some metastatic tumors of uncertain primary origin.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
N1NK
Concentration:
Greater than or equal to 69 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Del Sordo R et al. Histology and histopathology. 2014; 29(1):101-106
References 2:
Miettinen M et al. American Journal of Surgical Pathology. 2014; 38(3):410-420
References 3:
Paine SML et al. Neuropathology and applied neurobiology. 2014; 40:544-550
References 4:
Antic T et al. American Journal of Pathology. 2011; 136:872-880
Napsin A has a specific function in normal alveolar epithelium and is proposed to play a role in the proteolytic processing of surfactant precursors. Napsin A is reported to be predominantly expressed in lamellar bodies of type II pneumocytes, secondary lysosomes of alveolar macrophages, respiratory epithelium of terminal and respiratory bronchioles, plasma cells, within a subset of lymphocytes in normal lung, as well as in epithelial cells of renal tubules in normal kidney and is weakly expressed in normal spleen. Studies have reported that Napsin A is expressed in 90% of primary lung adenocarcinomas.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
IP64
Concentration:
Greater than or equal to 8.3 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Yamashita Y et al. Modern Pathology. 2015; 28: 111-11
References 2:
Kandalaft PL et al. American Journal of Clinical Pathology. 2014; 142: 830-836
Mismatch repair gene MutS Homolog 6 is a ubiquitous gene encoding the mismatch repair protein (MMR) MutS protein homolog 6 (MSH6). MSH6 functions by repairing mutations occurring during DNA replication, in normal proliferating cells.
Antibody Isotype:
IgG
Monosan Range:
MONXtra
Clone:
PU29
Concentration:
Greater than or equal to 200 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Warren J et al. Molecular Cell. 2007; 26:579-592
References 2:
Marti T et al. Journal of Cellular Physiology. 2002; 191:28-41.
Mismatch repair gene hMLH1 is a ubiquitous gene encoding the mismatch repair protein (MMR) MutL protein homolog 1 (MLH1). MLH1 functions by repairing mutations occurring during DNA replication, in normal proliferating cells.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
ES05
Concentration:
Greater than or equal to 165 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Tamura G et al.World Journal of Gastroenterology 2006; 12(2): 192198
References 2:
Abdel-Rahman W et al. Critical Reviews in Oncology/Hematology 2006; 58: 208220
References 3:
Mitchell R et al. American Journal of Epidemiology 2002; 156:885902
References 4:
Kuismanen S et al. American Journal of Pathology 2000; 156(5): 17731779
CD33 antigen is reported to appear on myelomonocytic precursor cells after CD34 antigen expression. It then continues to be expressed on both the myeloid and monocyte lineages, although it is reported to be absent on granulocytes. It has been reported that expression of CD33 is restricted to monocytes, premyelocytes, myeloid blasts, some acute undifferentiated leukemias and acute lymphoblastic leukemias.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
PWS44
Concentration:
Greater than or equal to 417 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Bradshaw EM et al. Nature Neuroscience. 2013, 16(7): 848 852
References 2:
Hoyer JD et al. American Journal of Clinical Pathology. 2008; 129(2): 316 323.
The CD7 molecule is a membrane-bound glycoprotein of 40 kD and is the earliest T cell specific antigen to be expressed in lymphocytes. CD7 antigen is also the only early marker to persist throughout differentiation. The function and role of the CD7 molecule has not yet been fully identified, although the activation of T cells with gamma/delta receptors has been proposed based on mAb-induced activation. CD7 antigen is reported to be found on the majority of peripheral blood T cells, most natural killer cells and thymocytes.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
LP15
Concentration:
Greater than or equal to 351 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Leong FJW et al. The Journal of Histotechnology. 2002; 25(4):215-227
References 2:
Ormsby A et al. Journal of the American Academy of Dermatology. 2001; 45(3):405-413
Geminin is a protein of 209 amino acids thought to be involved in the control of DNA replication via the interaction with Cdt1. Geminin is not found in the G1 phase of the cell cycle, but is first expressed in the G1 to S transition phase, with expression levels rising through the rest of the cell cycle and levels reaching a maximum during mitosis. It has been proposed that geminin may be a tumor suppressor protein. Geminin is reported to be expressed in proliferating lymphocytes and epithelial cells, for example, germinal centers in tonsil as well as in colon, spermatocytes, seminiferous tubules of the testes, within the basal layers of the squamous epithelium of the skin and breast. Geminin is reported to be upregulated in cancers such as non-Hodgkin's lymphoma, B cell lymphoma, breast carcinoma and colon carcinoma.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
EM6
Concentration:
Greater than or equal to 19 mg/L
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
McGarry TJ and Kirschner MW. Cell. 1998; 93:10431053
Prokaryotic recombinant protein corresponding to a 70 amino acid component of the N-terminal region of the cytokeratin 20 intermediate filament protein
Cytokeratin 20 has been demonstrated to be almost entirely confined to the gastric and intestinal epithelium, urothelium and Merkel cells of the skin. Cytokeratin 20 is less acidic than other type I cytokeratins and is of interest due to its restricted tissue expression. In normal tissue, cytokeratin 20 is expressed in intestinal epithelium, gastric foveolar epithelium, a number of endocrine cells in the upper portions of the pyloric glands, urothelium and Merkel cells in epidermis. In tumors it is reported, there is a marked difference in the expression of cytokeratin 20 within different carcinomas. Neoplasms expressing cytokeratin 20 are derived from normal epithelia which themselves expressed cytokeratin 20. Colorectal carcinomas consistently express cytokeratin 20, while gastric adenocarcinomas express cytokeratin 20 to a lesser degree. Adenocarcinomas of the gall bladder and bile duct, ductal cell adenocarcinomas of the pancreas, mucinous ovarian tumors, Merkel cell tumors and transitional cell carcinomas have also been reported to express cytokeratin 20.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
PW31
Concentration:
Greater than or equal to 37 mg/L
Storage buffer:
Tissue culture supernatant with 15 mM sodium azide
Storage:
2-8°C
References 1:
Campbell F and Herrington CS. Current Diagnostic Pathology. 2001; 7:113-122
References 2:
Leech SN et al. Journal of Clinical Pathology. 2001; 54:727-729
References 3:
Ferrari L et al. Anticancer Research. 1999; 19: 3415-3428
Calretinin is a calcium-binding protein of 29 kD that is a member of the family of so-called EF-hand proteins that also includes S-100 proteins. Calretinin is reported to be abundantly expressed in neurons. Outside the nervous system, calretinin is reported to be expressed in a range of cell types including mesothelial cells, steroid producing cells, (for example adrenal cortical cells, Leydig cells, ovarian theca interna cells, Sertoli cells, some neuroendocrine cells, eccrine sweat glands) and other cell types.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
CAL6
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Groves P et al. Acta Biochimica Polonica 2001, 48(1), 113-119
References 2:
Rogers JH .The Journal of Cell Biology 1987, 105, 1343-1353
Bcl-6 is a proto-oncogene that encodes a Kruppel-type zinc-finger protein of 95 kD and shares homology with other transcription factors. Bcl-6 protein is mainly expressed in normal germinal center B cells and related lymphomas. It has been shown that the Bcl-6 proto-oncogene is involved in chromosome rearrangements at 3q27 in non-Hodgkin's lymphomas and Bcl-6 rearrangements have also been detected in 33-45% of diffuse large B cell lymphomas. Immunohistochemistry has been reported to show the Bcl-6 gene product to be detectable in follicular lymphomas, diffuse large B cell lymphomas, Burkitt's lymphomas and in nodular, lymphocyte predominant Hodgkin's disease.
The CD3 molecule consists of five different polypeptide chains with molecular weights ranging from 16 to 28 kD. The CD3 antigen is first detected in early thymocytes and its appearance probably represents one of the earliest signs of commitment to the T cell lineage. Clone LN10 is specific for the non-glycosylated epsilon chain of the human CD3 molecule. Clone LN10 recognizes T cells in thymus, bone marrow, peripheral lymphoid tissue and blood and is a pan T cell marker.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
LN10
Concentration:
Greater than or equal to 32 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Krynitz B et al. Acta Dermato Venerologica. 2010; 90:379-385
Wilms' tumor protein (WT1) has a role in transcriptional regulation and is expressed in the kidney and a subset of hematopoietic cells. Alteration of transcription factor function is a common mechanism in oncogenesis. The WT1 protein contains a DNA binding domain and any deletions or point mutations of the WT1 gene which destroy this activity result in the development of the childhood nephroblastoma Wilms' tumor and Denys-Drash syndrome. The description of WT1 involvement in nephroblastoma is not clear.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
WT49
Concentration:
Greater than or equal to 44 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Omeroglu A and Omeroglu G. Arch Pathol Lab Med. 2003; 127:e347-e348
References 2:
Lee SB and Haber DA. Experimental Cell Research. 2001; 264:74-99
IL-6 is a multifunctional cytokine that is secreted by both lymphoid and non-lymphoid cells. It plays a key role in immune responses, hematopoiesis and is an important cytokine in cell proliferation and differentiation. It may also play an important role as an autocrine growth factor in metastatic prostate cancer. IL-6 has been reported to play a role in secretion or release of pituitary hormone in pituitary hormone secreting cells and adenomas. In addition, IL-6 has been suggested to have a trophic effect in nerve cells and to have a direct pathogenic role in CNS disorders. There are an increasing number of reports that cytokines of the IL-6 family play an important regulatory role in heart physiology.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
10C12
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
Salgado R et al. International Journal of Cancer. 103 (5): 642646 (2003)
References 2:
Kurotani R et al. Modern Pathology. 14 (8): 791797 (2001)
References 3:
Menet E et al. European Cytokine Network. 12 (4): 639646 (2001)
References 4:
Ono S et al. Journal of the Neurological Sciences. 187 (12): 2734 (2001)
References 5:
Yasukawa K et al. The EMBO Journal. 6 (10): 29392945 (1987)
Cytokeratins are intermediate filament proteins present in epithelial cells. They are expressed in a tissue-specific manner in normal organs and the tumors that arise from them. Cytokeratin 7 belongs to the neutral basic type B subfamily of cytokeratins. Its distribution is confined to glandular and transitional epithelia. Cytokeratin 7 is reported to be expressed in abundance in cultured bronchial and mesothelial cells but only at lower levels in cultured epidermal cells. The predicted amino acid sequence of this keratin has revealed a striking difference between this keratin and the type II keratins expressed in epidermal cells. Cytokeratin 7 has been reported in adenocarcinomas of the lung, breast, endometrium, ovary, thyroid as well as in carcinomas of the bladder and chromophobe renal cell carcinoma. Cytokeratin 7 and Cytokeratin 20 expression have been reported to show characteristic patterns on primary and metastatic lung and colorectal adenocarcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
RN7
Concentration:
Greater than or equal to 17 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
van de Molengraft FJJM et al.Histopathology. 1993; 22:35-38
References 2:
van Niekerk CC et al. Journal of Pathology. 1991; 165(2):145-15
CD79b, also known as B29 and Ig-beta is thought to function in the cellular activation and signaling that occurs when surface immunoglobulin (Ig) on B cells binds antigen or becomes cross-linked by anti-Ig antibody. This function occurs with the formation of a membrane signaling complex that is associated with Ig at the surface of B cells. CD79b, together with CD79a, forms the B cell antigen receptor (mlg) complex.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
JS01
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Matutes E. Journal of Clinical Pathology. 55: 180183 (2002)
References 2:
McCarron K F et al. American Journal of Clinical Pathology. 113 (6): 805813 (2000)
References 3:
Moreau E J et al. American Journal of Clinical Pathology. 108: 378382 (1997)
CD11c is a member of the leukocyte integrin family of adhesion proteins. It is reported to be expressed in normal tissues, mainly on myeloid cells, for example, in bone marrow myelocytes, premyelocytes, metamyelocytes, non-segmented and segmented neutrophils with high levels reported on tissue macrophages and monocytes and with lowest levels in granulocytes. It is also reported to be expressed on NK cells, activated T cells, lymphoid cell lines, including hairy cell leukemias and a proportion of interdigitating dendritic cells.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
5D11
Concentration:
Greater than or equal to 30 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Barth TFE et al. Journal of Pathology. 2007; 211:305-313
References 2:
Venkatraman L et al. Modern Pathology. 2005; 18: 255A-256A 1183 Supplement 1
aveolin-1 is a major structural component of caveolae, which are vesicular invaginations present on the plasma membrane of different cell types. It plays a regulatory role in several signaling pathways and is reported to be most abundantly expressed in terminally differentiated mesenchymal cells such as smooth muscle cells, adipocytes and endothelial cells. High levels are also reported in fibroblasts where a fine granular membranous and diffuse cytoplasmic staining pattern is described.
Antibody Isotype:
IgG2a, kappa
Monosan Range:
MONXtra
Clone:
4D6
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Wiechen K et al. American Journal of Pathology. 158 (3): 833839 (2001).
References 2:
Scherer PE et al. PNAS. 93: 131135 (1996)
References 3:
Lisanti MP et al. Journal of Cellular Biology. 126 (1): 111126 (1994).
Pax genes are a family of developmental control genes that encode nuclear transcription factors and have been implicated in the control of mammalian development. Pax-5 is a B cell specific transcription factor that is expressed in pro B cells, pre-B and mature B cells, and subsequently in all stages of B cell development until the plasma cell stage in which it is downregulated.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
1EW
Concentration:
Greater than or equal to 29 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Hansson M et al. European Journal of Haematology. 2007; 79:159-165
Human fascin is a 55 to 58 kD actin-bundling protein, whose actin binding ability is regulated by phosphorylation. In normal tissues the detection of fascin is reported to be predominantly restricted to dendritic cells, and in the thymus has been observed only in medullary dendritic cells. In reactive nodes, interdigitating reticulum cells of T cell zones, cells in subcapsular areas, and cells of the reticular network express fascin. Variable expression is seen in follicular dendritic cells and endothelial cells. Lymphoid cells, myeloid cells and plasma cells do not express fascin; however, in cases of Hodgkin's disease, including nodular sclerosis, mixed cellularity lymphocyte depletion and unclassified cases, most or all Reed Sternberg cells are reported to be positive for fascin. Fascin expression may be induced by Epstein-Barr virus (EBV) infection of B cells with the possibility that viral induction of fascin in lymphoid or other cell types must also be considered in EBV-positive cases.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
IM20
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15 mM Sodium azide
Storage:
2-8°C
References 1:
Ishikawa R et al. The Journal of Biological Chemistry. 273 (41): 2699126997 (1998)
References 2:
Ono S et al. The Journal of Biological Chemistry. 272 (4): 25272533 (1997)
References 3:
Pinkus GS et al. American Journal of Pathology. 150 (2): 543562 (1997)
E-cadherin is a Ca2+-dependent, transmembrane cell adhesion molecule. It plays an important role in the growth, development and the intercellular adhesion of epithelial cells. Most tumors have an abnormal architecture and any subsequent loss of adhesiveness is thought to be an important step in the development of local invasion. E-cadherin may have a role in neoplastic progression, particularly as a suppressor of invasion. In prostate cancers, for example, the expression of E-cadherin is reported to be reduced or absent in comparison with its expression in normal prostate which is uniformly strong. Reduced expression or absence of E-cadherin in addition to alpha, beta and gamma-catenin in primary breast carcinomas has also been reported and these four proteins are associated with the development of metastases.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
36B5
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Elston MS et al. J.of Clin.Endocrinology and Metabolism. 2009; 94(4):1436-1442.
References 2:
Munhoz NG et al. The Open Pathology Journal. 2009; 3:10-17
References 3:
Chetty R and Serra S. Histopathology 2008; 52: 325330
References 4:
Schott M et al. Endocrinology and Metabolism 2007; 92(9):3378- 3382
References 5:
Dansranjavin T et al. Oncology Reports. 2006; 15:1125-1131
The catenins, (alpha, beta and gamma) are cytoplasmic proteins which bind to the highly conserved tail of the E-cadherin molecule. Beta-catenin is a component of the adherens junction, a multiprotein complex which supports Ca2+ -dependent cell-to-cell contact, which in itself is critical for adhesion, signal transmission and for anchoring the actin cytoskeleton. Beta-catenin's role is as a transcription effector of the wnt-signaling pathway. Immunohistochemistry is the best way to demonstrate nuclear expression of beta-catenin and wnt-pathway activation. This aberrant expression is observed in human tumorigenesis, and especially in colorectal cancer.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
17C2
Concentration:
Greater than or equal to 51 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Curia MC et al. Modern Pathology. 2008; 21:7-14
References 2:
Ortega P et al. Clinical Cancer Research. 2008; 14(14):995-1001
References 3:
Daa T et al. J. of Exp.Clin.Cancer Research. 2005; 24(1):83-87
References 4:
Fadare O et al. World Journal of Surgical Oncology. 2005; 3(38)
References 5:
Gamachi A et al.Modern Pathology. 2003; 16(11):1124-1131
The neural cell adhesion molecules are a family of closely-related cell surface glycoproteins thought to play a role in embryogenesis, development and contact-mediated interactions between neural cells. The CD56 antigen (NCAM) consists of four major isoforms generated by differential splicing of the RNA transcript from a single gene located on chromosome 5. The CD56 antigen is expressed on neurons, astrocytes, Schwann cells, NK cells and a subset of activated T lymphocytes.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
CD564
Concentration:
Greater than or equal to 11 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Nakamoto Y et al. Clinical and Experimental Immunology. 2007; 147:296-305
References 2:
Wicherek L et al. Reproductive Biology and Endocrinology. 2006; 4:41
References 3:
Wicherek l et al. Journal of Reproductive Immunology. 2006; 70:119-131
The CD2 antigen (LFA-2) is a monomeric 45 to 58 kD glycoprotein. It is an accessory molecule important in mediating the adhesion of activated T cells and thymocytes with antigen-presenting cells and target cells.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
AB75
Concentration:
Greater than or equal to 56 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Leong FJW et al. The Journal of Histotechnology. 2002; 25(4):215227
Analyte Specific Reagent. Analytical and performance characteristics are not established. The product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
RBC2/3D5
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Analyte Specific Reagent. Analytical and performance characteristics are not established. Human gamma-sarcoglycan (35 kD). The product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG2b, kappa
Monosan Range:
MONXtra
Clone:
35DAG/21B5
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Analyte Specific Reagent. Analytical and performance characteristics are not established. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Human delta-sarcoglycan (35 kD). Does not react with delta-sarcoglycan in sections of mouse, rat, rabbit, dog, chicken, hamster or pig muscle. Other animal species not tested.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
dSarc3/12C1
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Human alpha-sarcoglycan, also known as adhalin. Also crossreacts strongly with alpha-sarcoglycan in sections of muscle from mouse, rat, rabbit, hamster and pig. Does not react with chicken muscle.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
Ad1/20A6
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003)
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Reconstitute with 1 mL or 0.1 mL of sterile distilled water as indicated on vial label. The myotilin gene on chromosome 5q31 encodes a 498 amino acid polypeptide with a molecular weight of 57kD. Myotilin is a structural protein of sarcomeric Z discs and sarcolemma in human skeletal and cardiac muscle. It is homologous to palladin and titin in the two C-terminal lg-domains and also to palladin in its unique serine-rich N-terminal region. Myotilin interacts with alpha-actinin, actin and gamma-filamin. Mutations in the myotilin gene are associated with limb-girdle muscular dystrophy 1 A (LGMD1A) and one form of Myofibrillar Myopathy. It is highly conserved between human and mouse with its expression being more widespread in the embryo than in the adult. Expression of myotilin has been reported in adult skeletal and cardiac muscle with variable expression reported in the peripheral nervous system, lung, liver and kidney. NCL-MYOTILIN will be of use in studies to determine the expression of myotilin in normal and pathological tissues.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
RSO34
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Mologni L et al. Mechanisms of Development. 103: 121125 (2001)
References 2:
Mykkänen OM et al. Mol. Biol. Cell. 12 (10): 30603073 (2001)
References 3:
Hauser MA et al. Human Molecular Genetics. 9 (14): 21412147 (2000)
References 4:
van der Ven PFM et al. Journal of Cell Biology. 151 (2): 235247 (2000)
References 5:
Salmikangas P et al. Human Molecular Genetics. 8 (7): 13291336 (1999
Myosin is a contractile muscle specific protein composed of two heavy and four light chains. The myosin heavy chain has many isoforms which are specific for different muscles or fiber types, some of which are developmentally regulated. The range of myosin heavy chain antibodies may prove useful for investigating development of intrafusal and extrafusal muscle fibers and the course of muscle fiber regeneration. At the ultrastructural level, antibodies can reveal architectural details of the myofilament as well as the cytoplasmic and membrane sites of new myosin integration. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Rabbit myosin slow type heavy chain. Crossreacts with human myosin slow type heavy chain.The antibody also reacts with type I myosin heavy chain in rat, mouse, dog, sheep, pig and goat muscle.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
WB-MHCS
Concentration:
n/a
Storage buffer:
Lyophilized tissue culture supernatant containing 15 mM sodium azide.
Storage:
2-8°C
References 1:
Sheriffs IN et al. Journal of Clinical Pathology. 54: 517520 (2001)
References 2:
Ecob-Prince M et al. Journal of Neurological Sciences. 91: 7178 (1989)
References 3:
Carson NE et al. The Journal of Histotechnology. 21 (1): 1924 (1998)
References 4:
Ecob-Prince M et al. Journal of Neurological Sciences. 90: 167177 (1989)
References 5:
Vivarelli E et al. Journal of Cellular Biology. 107: 21912197 (1988)
Myosin is a contractile muscle specific protein composed of two heavy and four light chains. The myosin heavy chain has many isoforms which are specific for different muscles or fiber types, some of which are developmentally regulated. The range of myosin heavy chain antibodies may prove useful for investigating development of intrafusal and extrafusal muscle fibers and the course of muscle fiber regeneration. At the ultrastructural level, antibodies can reveal architectural details of the myofilament as well as the cytoplasmic and membrane sites of new myosin integration. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Rabbit myosin fast type heavy chain. Crossreacts with human myosin fast type heavy chain. Rabbit myosin neonatal type heavy chain. Crossreacts with human myosin neonatal type heavy chain. Note that this antibody recognises a myosin heavy chain present during the neonatal period in rabbit limb muscle. The temporal appearance of an equivalent epitope may differ in different species and consequently it may not be correct to label the epitope as neonatal in some circumstances.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
WB-MHCN
Concentration:
n/a
Storage buffer:
Lyophilized tissue culture supernatant containing 15 mM sodium azide.
Storage:
2-8°C
References 1:
Ecob-Prince M et al. Journal of Neurological Sciences. 90: 167177 (1989)
References 2:
Ecob-Prince M et al. Journal of Neurological Sciences. 91: 7178 (1989)
Myosin is a contractile muscle specific protein composed of two heavy and four light chains. The myosin heavy chain has many isoforms which are specific for different muscles or fiber types, some of which are developmentally regulated. The range of myosin heavy chain antibodies may prove useful for investigating development of intrafusal and extrafusal muscle fibers and the course of muscle fiber regeneration. At the ultrastructural level, antibodies can reveal architectural details of the myofilament as well as the cytoplasmic and membrane sites of new myosin integration. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Rabbit myosin fast type heavy chain. Crossreacts with human myosin fast type heavy chain. The antibody also reacts with type II myosin heavy chain (both IIa and IIb) in rat, mouse, dog, sheep, pig and goat muscle.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
WB-MHCf
Concentration:
n/a
Storage buffer:
Lyophilized tissue culture supernatant containing 15 mM sodium azide.
Storage:
2-8°C
References 1:
Sheriffs IN et al. Journal of Clinical Pathology. 54: 517520 (2001)
References 2:
Ecob-Prince M et al. Journal of Neurological Sciences. 91: 7178 (1989)
References 3:
Carson NE et al. The Journal of Histotechnology. 21 (1): 1924 (1998)
References 4:
Hoh JFY and Hughes A. Journal of Muscle Research and Cellular Motility. 10: 312325 (1989)
Myosin is a contractile muscle specific protein composed of two heavy and four light chains. The myosin heavy chain has many isoforms which are specific for different muscles or fiber types, some of which are developmentally regulated. The range of myosin heavy chain antibodies may prove useful for investigating development of intrafusal and extrafusal muscle fibers and the course of muscle fiber regeneration. At the ultrastructural level, antibodies can reveal architectural details of the myofilament as well as the cytoplasmic and membrane sites of new myosin integration. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
RNMy2/9D2
Concentration:
n/a
Storage buffer:
Lyophilized tissue culture supernatant containing 15 mM sodium azide.
Storage:
2-8°C
References 1:
Davis CE et al. Neuromuscular Disorders. 1 (6): 411421 (1991)
References 2:
Ecob-Prince M et al. Journal of Neurological Sciences. 91: 7178 (1989)
The muscle-specific form of laminin, merosin, is composed of three chains: alpha 2, beta 1 and gamma 1.Analyte Specific Reagent. Analytical and performance characteristics are not established. Reacts strongly with laminin alpha 2 chain of merosin in human and rabbit skeletal muscle. No reaction is observed in muscle sections from mouse, rat, dog, chicken, hamster or pig. The product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
Mer3/22B2
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Analyte Specific Reagent. Analytical and performance characteristics are not established. NCL-EMERIN is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
4G5
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Muschen M et al. Cancer Research. 61 (5): 20802084 (2001)
References 2:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Dystrophin is the 427kD protein product of the DMD gene located on the X chromosome at position Xp21. Analyte Specific Reagent. Analytical and performance characteristics are not established. Product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
34C5
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Blake DJ et al. Physiological Reviews. 82 (2): 291329 (2002)
References 2:
Oliveira AS et al. Arq Neuropsiquiatr. 50 (4): 478485 (1992
References 3:
Haginoya K et al. Journal of Neurology. 238 (7): 375378 (1991)
Dystrophin is the 427kD protein product of the DMD gene located on the X chromosome at position Xp21. Analyte Specific Reagent. Analytical and performance characteristics are not established. Product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Reacts strongly with the amino terminal domain (between amino acids 321 and 494) of human dystrophin. Patient immunoreactivity indicates epitope is near exons 10 to 12. Epitope mapping suggests that sequences from amino acids 308 to 351 are involved in antibody binding. This region spans the junction of exons 9 and 10 and the epitope recognised may be part of a hinge region joining the amino domain to the central rod domain. No reactivity with DMD/BMD patients deleted for exons 10 to 12. No crossreaction is observed with mouse (high background only), rat, rabbit, dog, chicken, hamster and pig dystrophin.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
DY10/12B2
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Dystrophin is the 427kD protein product of the DMD gene located on the X chromosome at position Xp21. Analyte Specific Reagent. Analytical and performance characteristics are not established. Product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Reacts strongly with the carboxy terminus (between amino acids 3669 and 3685) of human dystrophin. Also crosssreacts strongly with skeletal, cardiac and smooth muscle dystrophin from normal mouse, rat, rabbit, dog, chicken and hamster. No crossreactivity with mdx mouse tissue. Crossreacts very weakly with pig dystrophin.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
DY8/6C5
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Dystrophin is the 427kD protein product of the DMD gene located on the X chromosome at position Xp21. Analyte Specific Reagent. Analytical and performance characteristics are not established. Product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label. Reacts strongly with the rod domain (between amino acids 1181 and 1388) of human dystrophin. Also reacts with skeletal, cardiac and smooth muscle dystrophin from normal mouse, rat, rabbit, dog, hamster and pig. No reactivity with mdx mouse tissue of DMD/BMD patients who have a gene deletion which removes the antibody binding site. No reaction is seen with chicken dystrophin.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
Dy4/6D3
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Analyte Specific Reagent. Analytical and performance characteristics are not established. The product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
Ham3/17B2
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Analyte Specific Reagent. Analytical and performance characteristics are not established. The product is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
Ham1/7B6
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Marafioti T et al. American Journal of Pathology. 162 (3): 861871 (2003
References 2:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 3:
Re D et al. Cancer Research. 61 (5): 20802084 (2001)
References 4:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
This antibody reacts with full-size calpain 3 (94 kD) plus an additional breakdown product at 60 kD in human skeletal muscle. The 94 kD band can be seen in muscle extracts from rabbit, mouse, dog, chicken, hamster, pig and rat. Degraded calpain 3 bands starting at approximately 60 kD are also usually present. Additional bands corresponding in size to calpains 1 and/or 2 can be detected in skeletal muscle from mouse, rat, chicken and hamster. Reconstitute with the volume of sterile distilled water indicated on the vial label.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
Calp3c/12A2
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Anderson LVB et al. American Journal of Pathology. 153 (4): 11691179 (1998)
References 2:
Topaloglu H et al. Neuropediatrics. 28: 212216 (1997)
This antibody reacts with full-size calpain 3 (94 kD) plus an additional fragment at 30 kD in human skeletal muscle. A band of 94 kD is seen with rabbit and dog muscle while extracts of hamster muscle show reactivity with the 94 kD band and a larger species of approximately 110 kD. This 110 kD band is the principal immunoreactive species seen in rat muscle extracts. This antibody produces no bands with mouse, pig or chicken muscle. Reconstitute with the volume of sterile distilled water indicated on the vial label.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
Calp3d/2C4
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Anderson LVB et al. American Journal of Pathology. 153 (4): 11691179 (1998)
References 2:
Topaloglu H et al. Neuropediatrics. 28: 212216 (1997)
NCL-b-DG is a lyophilized tissue culture supernatant containing sodium azide as a preservative. The user is required to reconstitute the contents of the vial with the correct volume of sterile distilled water as indicated on the vial label.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
43DAG1/8D5
Concentration:
n/a
Storage buffer:
Lyophilized
Storage:
2-8°C
References 1:
Hess J et al. Molecular and Cellular Biology. 21 (5): 15311539 (2001)
References 2:
Muschen M et al. Cancer Research. 61 (5): 20802084 (2001)
References 3:
Luo Y and Roeder R G. Molecular and Cellular Biology. 15 (8): 41154124 (1995)
Synaptophysin is an integral membrane glycoprotein with a molecular weight of 38 kD. It is reported to occur in presynaptic vesicles of neurons in brain, spinal cord, retina, in similar vesicles of the adrenal medulla as well as in neuromuscular junctions. Synaptophysin may be involved in synaptic vesicle formation and exocytosis. Synaptophysin is reported to be expressed in a wide spectrum of neuroendocrine tumors including neuroblastomas, ganglioneuroblastomas, phaeochromocytomas, chromaffin and non-chromaffin paragangliomas. Synaptophysin is also reported to be expressed in neuroendocrine tumors of epithelial type including pituitary adenomas, islet cell tumors, medullary carcinomas of thyroid, parathyroid adenomas, carcinoids of the bronchopulmonary and gastrointestinal tracts, neuroendocrine carcinomas of the bronchopulmonary and gastrointestinal tract and neuronendocrine carcinomas of the skin.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
27G12
Concentration:
Greater than or equal to 42 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Takeda S et al. Neuropathology. 2003; 23(4):254261
Protein gene product (PGP) 9.5 is a neuron-specific protein, structurally and immunologically distinct from neuron specific enolase. The protein which has a molecular weight of 27 kD was first identified by high resolution two dimensional PAGE. PGP9.5 expression has been reported in neurons and nerve fibers at all levels of the central and peripheral nervous system, in many neuroendocrine cells, in segments of the renal tubules, in spermatogonia and Leydig cells of the testis, in ova and in some cells of both the pregnant and non-pregnant corpus luteum. PGP9.5 is a member of the ubiquitin C-terminal hydroxylase family and is also concentrated within inclusion bodies suggesting that such structures may be metabolically active regions of the cells.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
10A1
Concentration:
Greater than or equal to 35 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Beresford L et al. Immunology. 2004; 111(1):118125
Neurofilaments constitute the main structural elements of neuronal axons and dendrites. Neurofilaments are composed of three major subunits referred to as the neurofilament triplet, with molecular weights of 68 kD, 160kD and 200 kD. Within tumors, only neoplastic cells of neural origin or those exhibiting neuronal differentiation, have been reported to express neurofilaments. NF-H (200 kD) polypeptide of human neurofilament. This antibody reacts with both phosphorylated and unphosphorylated forms of NF-H.
Alpha-synuclein is a protein of 140 amino acids and a member of the synuclein family. It shares 61% sequence homology with beta-synuclein and is highly conserved between vertebrate species. It does not possess a signal sequence suggesting that it is an intracellular protein. All synucleins have an unusual organization based around the eleven residue repeating motif and an alpha-helical secondary structure resembling those found in the lipid-binding domain of exchangeable apolipoproteins, including Apo E. This homology suggests a direct interaction of alpha-synuclein with membranes consistent with its affinity for synaptosomes.Clone KM51 is specific for alpha-synuclein and is unreactive with beta-synuclein. Pretreatment of tissue sections with 98-100% formic acid is also recommended.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
KM51
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Baba M et al. American Journal of Pathology. 152 (4): 879884 (1998)
References 2:
Polymeropoulos MH et al. Science. 276: 20452047 (1997)
References 3:
Spillantini MG et al. Nature. 388: 839840 (1997)
References 4:
Weinreb PH et al. Biochemistry. 35 (43): 1370913715 (1996)
NCL-CK5/6/8/18, NCL-L-CK5/6/8/18 and RTU-CK5/6/8/18 react with human cytokeratins 5, 6, 8 and 18. These products are cocktails of monoclonal antibodies designed to recognize cytokeratins reported to be expressed in almost all epithelial tissues.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
5D3/LP34
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15 mMsodium azide
Storage:
2-8°C
References 1:
Hatzfeld M and Weber K. Journal of Cell Biology. 116: 157166 (1992)
References 2:
Angus B et al. Journal of Pathology. 155: 7175 (1988)
References 3:
Ghosh A K et al. British Journal of Haematology. 61: 2130 (1985)
References 4:
Gatter KC et al. Journal of Clinical Pathology. 35: 12531267 (1982)
Clones AE1 and AE3 are specific for the 56.5, 50, 50', 48 and 40 kD acidic cytokeratins as well as the 65 to 67, 64, 59, 58, 56 and 52 kD basic cytokeratins. The cocktail of clones AE1 and AE3 exhibit broad reactivity with two families of cytokeratin, acidic and basic.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
AE1/AE3
Concentration:
Greater than or equal to 225 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Nadji M, Morales AR. Laboratory Medicine. 1983; 14:767
References 2:
Omata M et al. Am.J.of Clin. Pathol. 1980; 73:626
References 3:
Su T et al. Diagnostic Pathology. 2014; 9: 179
References 4:
Zhao W et al. Int.J. of Clin. and Exp.Pathol. 2014; 7(11): 7951-7956
References 5:
Hammers HJ et al. Molecular Cancer Therapeutics. 2010; 9(6): 1525-1535
Cytokeratins 5, 6 and 18 are reported to be expressed in a broad range of human epithelial tissues, from simple glandular epithelia to stratified squamous epithelia. These include epithelial cells that are ectodermal, mesodermal, or endodermal in origin. These cytokeratins have been reported to be expressed in tumor cells of epithelial origin and less commonly of mesothelial origin. Non-epithelial tumors such as lymphomas do not express these cytokeratins. The recognition of cytokeratin 18 on formalin fixed paraffin embedded sections using clone LP34 may be variable.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
LP34
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Lyall F et al. The American Journal of Pathology. 2001;159(5):1827-1838
References 2:
Yokozaki H et al. Japanese Journal of Clinical Oncology. 2000;30(2):101-104
References 3:
Lyall F et al. The American Journal of Pathology. 1999;154(4):1105-1114
Clone 5D3 reacts with human cytokeratin intermediate filament proteins of 52.5 kD and 45 kD, identified as cytokeratins 8 and 18, respectively. Clone 5D3 shares similar specificities to clone CAM5.2 (Angus B et al. Journal of Pathology. 153: 377-384 (1987)).
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
5D3
Concentration:
Greater than or equal to 420 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Nadji M, Morales AR. Laboratory Medicine. 1983; 14:767
References 2:
Omata M et al. Am.J.of Clin. Pathol. 1980; 73:626
References 3:
Angus B et al. J.of Pathology. 1988; 155(1):7175
References 4:
Martin CA et al. Appl Immunohistochem Mol Morphol. 2001; 9(1):7073
References 5:
Mehes G et al. Am. J.of Pathol. 2001; 159(1):1720
Cytokeratin 20 has been demonstrated to be almost entirely confined to the gastric and intestinal epithelium, urothelium and Merkel cells of the skin. Cytokeratin 20 is less acidic than other type I cytokeratins and is of interest due to its restricted tissue expression. In normal tissue, cytokeratin 20 is expressed in intestinal epithelium, gastric foveolar epithelium, a number of endocrine cells in the upper portions of the pyloric glands, urothelium and Merkel cells in epidermis. In tumors it is reported, there is a marked difference in the expression of cytokeratin 20 within different carcinomas. Neoplasms expressing cytokeratin 20 are derived from normal epithelia which themselves expressed cytokeratin 20. Colorectal carcinomas consistently express cytokeratin 20, while gastric adenocarcinomas express cytokeratin 20 to a lesser degree. Adenocarcinomas of the gall bladder and bile duct, ductal cell adenocarcinomas of the pancreas, mucinous ovarian tumors, Merkel cell tumors and transitional cell carcinomas have also been reported to express cytokeratin 20.
Antibody Isotype:
IgG2a, kappa
Monosan Range:
MONXtra
Clone:
Ks20.8
Concentration:
Greater than or equal to 9,26 mg/L
Storage buffer:
PBS (pH 7.6) with 1% BSA and sodium azide
Storage:
2-8°C
References 1:
Botta MC et al. Pathologica. 2001; 93(6):640-644
References 2:
Leech SN et al. Journal of Clinical Pathology. 2001; 54:727-729
References 3:
Tan J et al. Human Pathology. 1998; 29(4):390-396
References 4:
Longatto Filho A et al. Acta Cytol. 1997; 41(4):961-971
References 5:
Moll R et al. American Journal of Pathology. 1992; 140(2):427-447
Cytokeratins 14 and 5 are useful to distinguish stratified epithelial cell types from simple epithelial cell types. Cytokeratin 14 has been reported to be expressed in neoplasms of squamous cell origin.Clone LL002 reacts with the human cytokeratin intermediate filament protein (50 kD) identified as cytokeratin 14.
Antibody Isotype:
IgG3
Monosan Range:
MONXtra
Clone:
LL002
Concentration:
Greater than or equal to 24 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Reis-Filho JS et al. Journal of Clinical Pathology. 2004; 57:83-86
References 2:
Sivard P et al. Experimental Dermatology. 2003; 12(4):346355
References 3:
Fong LYY et al. Cancer Research. 2003; 63:186195
References 4:
Nagao T et al. Histopathology. 2001; 38(1):3036
References 5:
Nakayama H et al. Japanese Journal of Clinical Oncology. 1997; 27(6):427432
Cytokeratins are intermediate filament proteins present in epithelial cells. They are expressed in a tissue-specific manner in normal organs and the tumors that arise from them. Cytokeratin 7 belongs to the neutral basic type B subfamily of cytokeratins. Its distribution is confined to glandular and transitional epithelia. Cytokeratin 7 is reported to be expressed in abundance in cultured bronchial and mesothelial cells but only at lower levels in cultured epidermal cells. The predicted amino acid sequence of this keratin has revealed a striking difference between this keratin and the type II keratins expressed in epidermal cells. Cytokeratin 7 has been reported in adenocarcinomas of the lung, breast, endometrium, ovary, thyroid as well as in carcinomas of the bladder and chromophobe renal cell carcinoma. Cytokeratin 7 and Cytokeratin 20 expression have been reported to show characteristic patterns on primary and metastatic lung and colorectal adenocarcinomas.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
OV-TL 12/30
Concentration:
Greater than or equal to 67 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
van de Molengraft FJJM et al.Histopathology. 1993; 22:35-38
References 2:
van Niekerk CC et al. Journal of Pathology. 1991; 165(2):145-15
Cytokeratins are a large family of cytoskeletal proteins found in epithelial cells. They are co-ordinately synthesized in pairs so that at least one member of each family is expressed in each epithelial cell. Cytokeratins assemble into obligatory heteropolymers composed of type I (acidic) and type II (basic) polypeptides to form higher order tetramers and protofilaments. Basal cells of human epidermis express acidic keratin 14 and basic cytokeratin 5. Cytokeratin 5 is a 58 kD protein that is closely related to cytokeratin 6. Point mutations in the cytokeratin 5 gene at locus 12q11-q13 can cause various types of epidermolysis bullosa simplex. Cytokeratin 5 is also reported to be expressed in most epithelial and biphasic mesotheliomas. Clone XM26 is specific for the 58 kD intermediate filament protein known as cytokeratin 5. It is not cross-reactive with cytokeratin 6.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
XM26
Concentration:
Greater than or equal to 21 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Bhargava R et al. The American Journal of Clinical Pathology . 2008; 130:724-730
References 2:
Laakso M e al. Clinical Cancer Research. 2006; 12(14):4185-4191
References 3:
Miettinen M et al. American Journal of Surgical Pathology. 2003; 27(2):150158
References 4:
Zhang RR et al. Breast Cancer Research. 2003; 5:R151R156
DES-DERII reacts with an 18 kD rod piece of the intermediate filament protein desmin (53 kD) in muscle cells. The antibody does not appear to recognize other intermediate filament proteins. In normal tissues, Clone DE-R-11 reacts with both striated (skeletal and cardiac) and smooth muscle cells. The labeling is confined to the Z bands in skeletal and cardiac muscle giving a characteristic striated appearance.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
DE-R-11
Concentration:
Greater than or equal to 27 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Beaton LJ et al. Journal of Physiology. 2002; 544(Pt3):849-859
References 2:
Barber A et al.The FASEB Journal. 2001; 15:1158-1168
References 3:
Debus E et al.The EMBO Journal. 1983; 2(12):2305-2312
References 4:
Baghdiguian S et al. Nature Medicine. 1999; 5(5):503-511
References 5:
Lyall F et al. American Journal of Pathology. 2001; 159(5):1827-1838
Cytoplasmic actins are part of the microfilament system of cytoskeletal proteins. Smooth muscle actin is found in vascular walls, intestinal muscularis mucosae and muscularis propria and in the stroma of various tissues. Enzyme pretreatment may enhance staining in some cases. Human alpha smooth muscle actin. Reactive with smooth muscle cells in blood vessel walls, gut wall, myometrium and arrectores pili of skin. Myoepithelial cells such as those in breast and salivary gland also contain actin.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
alpha sm-1
Concentration:
Greater than or equal to 4,5 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Skalli O et al. The Journal of Cell Biology. 1986; 103:2787-2796
References 2:
Ramos JG et al. Endocrinology. 2003; 144(7):3206-3215
References 3:
Suárez-Vilela D and Izquierdo-Garcia FM. Histopathology. 2003; 43(4):398-400
References 4:
Vicario JH et al. Rev Fed Arg Cardiol. 2002; 31:441-449
References 5:
Barry-Lane PA et al. Journal of Clinical Investigation. 2001; 108(10):1513-1522
Terminal deoxynucleotidyl transferase (TdT) is a DNA polymerase of 58 kD located in the cell nucleus which catalyzes the polymerization of deoxynucleotides at the 3' hydroxyl ends of oligo or polydeoxynucleotide initiators and functions without a template. TdT is reported to be expressed in primitive T and B lymphocytes of the normal thymus and bone marrow. The identification of TdT-positive cell populations in primary and secondary lymphoid organs during maturation of the immune system is one area of interest but it is the reported occurrence of high levels of enzyme activity in white blood cells and bone marrow in certain leukemias which is of particular interest.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
SEN28
Concentration:
Greater than or equal to 30 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Tai YC and Peh SC. FSingapore Medical Journal. 2003; 44(5):250-255
Prokaryotic recombinant protein corresponding to a region which spans the tyrosine kinase catalytic domain and part of the C-terminus of the NPM-ALK transcript (419-520??).
Anaplastic large cell lymphoma (ALCL) is usually composed of large pleomorphic cells which are reported to express CD30 antigen and epithelial membrane antigen (EMA). These tumor cells tend to occur in younger individuals and may be associated with cutaneous and extranodal involvement. A proportion of these cases contain a chromosomal translocation t(2;5) (p23;q35). This results in a hybrid gene encoding part of the nucleophosmin (NPM) gene joined to the cytoplasmic domain of the anaplastic lymphoma kinase (ALK) gene, giving rise to the protein, p80. Large cell lymphomas account for approximately 25% of all non-Hodgkin's lymphomas in children and young adults, of which one third carry the NPM-ALK gene translocation.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
5A4
Concentration:
Greater than or equal to 32 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Liu A et al. Acta Histochemica et Cytochemica. 2004; 37(1):21-30
Granzymes are neutral serine proteases which are stored in specialized lytic granules of cytotoxic T lymphocytes (CTL) and in natural killer (NK) cells. These CTL and NK cells are heavily involved in the elimination of neoplastic and virally infected cells. Secretory granules containing perforin and granzymes are instrumental in undertaking cytolytic activity. Granzyme B is understood to enter a target cell through a perforin pore-formed channel to induce DNA fragmentation and apoptosis. Granzyme B has also been described in neoplastic CTL and NK cells.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
11F1
Concentration:
Greater than or equal to 47 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Lee HK et al. J Korean Med Sci. 2003; 18:272276
References 2:
Theate I et al.European Journal of Haematology. 2002; 69(4):248253
Human von Willebrand factor (or factor VIII-related antigen) is a 270 kD multimeric plasma glycoprotein. It mediates platelet adhesion to injured vessel walls and serves as a carrier and stabilizer for coagulation factor VIII. The von Willebrand factor has functional binding domains to platelet glycoprotein Ib, glycoprotein Ib/IIIa, collagen and heparin. Von Willebrand factor is synthesized by endothelial cells and is reported to be expressed in a number of tumors of vascular origin.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
36B11
Concentration:
Greater than or equal to 21 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Zhang Y et al. Human Pathology. 2005; 36:797-805
References 2:
Su H et al. Proceedings of the National Academy of Sciences, USA. 2000; 97(25):13801-13806
The biosynthesis of melanin in melanocytes involves a family of enzymes, a key member of which is tyrosinase. Tyrosinase deficiency is associated with various forms of albinism and in particular oculocutaneous albinism. L-tyrosinase is the initial substrate for melanin biosynthesis and its conversion to dopaquinone is catalyzed by tyrosinase, whose expression is reported in melanocytes and melanomas.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
T311
Concentration:
Greater than or equal to 89 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Shidham VB et al. BMC Cancer. 2003; 3(1):15
References 2:
Lohmann CM et al. American Journal of Surgical Pathology. 2002; 26(10):13511357
References 3:
Clarkson KS et al. Journal of Clinical Pathology. 2001; 54(3):196200
References 4:
de Vries TJ et al. Journal of Pathology. 2001; 193(1):1320
References 5:
Jungbluth AA et al. Pathol Res Pract. 2000; 196(4):235242
Thyroid Transcription Factor-1 (TTF-1) is a member of the homeodomain transcription factor family and plays a role in regulating genes expressed within the thyroid, lung and brain. These include thyroglobulin, thyroid peroxidase, Clara cell secretory protein and surfactant proteins. Human TTF-1 (38 kD) is a single polypeptide of 371 amino acids sharing 98% homology with the equivalent rat and mouse proteins. TTF-1 functions by binding to specific recognition sites in a manner that may be regulated by both the redox and phosphorylation status of the protein. In addition to its role as a tissue-specific transcriptional activator in adult organs, TTF-1 may also function in organogenesis. Gene targeting studies have shown TTF-1 to be essential for the proper development of the thyroid and lungs and abnormal expression may underline a number of congenital abnormalities.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
SPT24
Concentration:
Greater than or equal to 108 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Unal B et al. Turkish Journal of Pathology. 2014; 30(3): 201-205
References 2:
Klingen TA et al. Diagnostic Pathology. 2013; 8: 80-86
References 3:
Berghmans T et al. Lung Cancer. 2006; 52(2): 219-224
References 4:
Penman D et al. Journal of Clinical Pathology. 2006; 59:663-664.
References 5:
Comperat E et al. Modern Pathology. 2005; 18(10):1371-1376
Prostatic acid phosphatase (PAP) is an isoenzyme of acid phosphatase found in large amounts in the prostate and seminal fluid. The precise function of PAP is unknown, but it may act as a hydrolase to split phosphoryl choline in semen and also function as a transferase. Elevated serum levels of the enzyme are reported in metastatic prostatic carcinoma.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
PASE/4LJ
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Haines AMR et al. British Journal of Cancer. 60: 887892 (1989)
References 2:
Haines AMR et al. Biochemical Society Transactions. 15: 11791180 (1987)
Prostate specific antigen (PSA) is a 34 kD protein belonging to the kallikrein family of serine proteases and was originally isolated and purified from human seminal plasma. It was found to be immunologically identical and biologically similar to a protein isolated from the prostate gland. PSA is distinct from prostatic acid phosphatase. Low levels of expression of PSA have been reported in non-prostatic tissues and tumors such as breast carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
35H9
Concentration:
Greater than or equal to 43 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Watt KWK et al. Proceedings of the National Academy of Sciences USA. 1986; 83:3166-3170
Placental alkaline phosphatase (PLAP) is a membrane-associated sialoglycoprotein enzyme normally present at high concentration in syncytiotrophoblasts within the placenta during the third trimester of gestation. The expression of PLAP was originally thought to be restricted to term placenta but a human PLAP-like variant has been described which shares more than 85% homology with PLAP itself. This high degree of homology between PLAP and PLAP-like enzyme together with cross-reacting antibodies has led to some confusion of the distribution of PLAP and PLAP-like enzyme in various tissues. PLAP is reported to be expressed only in normal term placenta, endocervix and fallopian tube and also in ovarian and proximal gastrointestinal tumors. PLAP expression is rare in malignant germ cell tumors. PLAP-like enzyme is reported to be predominantly found in normal fetal and neonatal testis, and in thymus. It is also commonly expressed in germ cell tumors and more recently described in seminomas.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
8A9
Concentration:
Greater than or equal to 26 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Bartkova J et al. Oncogene. 2000; 19: 4146-4150
References 2:
Franke FE et al. Human Pathology 2000; 31(12), 14661476
References 3:
Hoei-Hansen CE et al. Molecular Cancer. 2007; 6:12
References 4:
McCann-Crosby B et al. International Journal of Pediatric Endocrinology 2015; 1:14
Rhabdomyosarcomas are a class of myoblast-derived soft tissue sarcomas that usually express a number of muscle-specific genes and primarily affect children and young adults. Differentiation of myogenic cells is controlled by a set of regulatory genes including MyoD1, myogenin, Myf-5 and Myf-6. Myf-4 is the human homolog of myogenin. Its gene product, together with that of Myf-3, accumulates in the nucleus of differentiated cells.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
LO26
Concentration:
Greater than or equal to 13 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Den Bakker MA et al. Histopathology. 2003; 43(3):297299
References 2:
Ingeholm P et al. APMIS. 2002; 110(9):639645
References 3:
Kumar S et al. Modern Pathology. 2000; 13(9):988993
References 4:
Gilpin BJ et al. Journal of Biological Chemistry. 1998; 273(1):157166
Microphthalmia transcription factor (MITF) gene product, a nuclear transcription factor of the basic-helix-loop-helix type, is thought to play a role in the regulation of genes encoding the enzymes necessary for melanogenesis. These include tyrosinase, TRP-1 and TRP-2. MITF is critical for the embryonic development and postnatal viability of melanocytes. The melanocyte-specific isoform of microphthalmia transcription factor MITF-M, is reported to be expressed in normal and malignant melanocytes. The other isoforms, MITF-A, MITF-C and MITF-H, differ structurally at the N-terminus from MITF-M.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
34CA5
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM Sodium azide
Storage:
2-8°C
References 1:
Fang D and Setaluri V. Biochem. and Biophys. Research Comm. 256 (3): 657663 (1999)
References 2:
King R et al. American Journal of Pathology. 155 (3): 731738 (1999)
References 3:
Amae S et al. Biochem.and Biophys.Research Comm. 247: 710715 (1998)
References 4:
Watanabe A et al. Nature Genetics. 18: 283286 (1998)
Recombinant prokaryotic fusion protein corresponding to approximately 100 amino acids which are present in the membrane-bound form of the mesothelin molecule.
Mesothelin is a glycosyl-phosphatidylinositol-linked (GPI) glycoprotein of 40kD present on the surface of mesothelial cells, mesotheliomas, epithelial ovarian cancers and some squamous cell carcinomas. It is synthesized as a 69 kD precursor which is enzymatically processed into an N-terminal secreted form of 30 kD and the GPI-linked membrane-bound form of 40 kD. The secreted form is identical to the megakaryocyte potentiating factor, but it is the GPI-linked membrane-bound form which has generated interest. Mesothelin is abundantly expressed in the kidney and in occasional epithelial cells of the trachea, tonsil and fallopian tube. The function of mesothelin is unclear but it may have a role in cellular adhesion. Mesothelin is reported to be abundant in the normal mesothelial cells from which malignant mesotheliomas and ovarian cystadenocarcinomas are derived.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
5B2
Concentration:
Greater than or equal to 40 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Ordonez NG. American Journal of Surgical Pathology. 2003; 27(11):14181428
References 2:
Ordonez NG. Modern Pathology. 2003; 16(3):192197
References 3:
Argani P et al. Clinical Cancer Research. 2001; 7(12):38623868
Melan A, a product of the MART-1 gene, is a melanocyte differentiation marker recognized by autologous cytotoxic T lymphocytes. Other melanoma-associated markers recognized by autologous cytotoxic T cells are reported to include MAGE-1, MAGE-3, tyrosinase, gp100, gp75, BAGE-1 and GAGE-1. The analysis of these different molecules and their expression in individual melanomas may be of help in the study of their particular molecular roles in melanocyte differentiation and tumorigenesis.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
A103
Concentration:
Greater than or equal to 22 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Shidham VB et al. BioMed Central cancer. 2003; 3(1):15
References 2:
Clarkson KS et al. Journal of Clinical Pathology. 2001; 54:196200
References 3:
De Vries TJ et al. Journal of Pathology. 2001; 193:1320
References 4:
Fang D et al. American Journal of Pathology. 2001; 158(6):21072115
References 5:
Chen YT et al. Proceedings of the National Academy of Sciences USA. 1996; 93:59155919
Gross cystic disease of the breast is a benign premenopausal disorder in which cysts are a predominant pathological lesion. These cysts appear to be formed from excessive apocrine cystic secretions. This fluid is composed of several glycoproteins including a unique 15 kD monomer protein, GCDFP15. It has been reported that cytosolic analysis of normal tissue from all major organs has demonstrated GCDFP15 in apocrine epithelia, lacrimal, ceruminous and Moll's glands and in numerous serous cells of the submandibular, tracheal, bronchial, sublingual and minor salivary glands. Specificity Human gross cystic disease fluid protein (15 kD)
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
23A3
Concentration:
Greater than or equal to 55 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Sapino A et al. J. of Biol.Regulators & Homeostatic Agents. 2000; 14(4):259262
References 2:
Haagensen DE Jr et al. Annals of the New York Academy of Sciences.1990;586:161173
Epithelial membrane antigen (EMA), also known as episialin, is reported to be expressed in a variety of normal and neoplastic epithelia. It has been reported that markers to CD45 (LCA) when used in conjunction with markers to EMA are useful in labeling cells of lymphoid origin, whereas the combination of anti-cytokeratin antibodies together with EMA is useful to characterize cells of epithelial origin. EMA is also notably described to be expressed in a subset of Hodgkin's lymphomas.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
GP1.4
Concentration:
Greater than or equal to 11 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Kim JH et al. The Korean Journal of Pathology. 2002; 36:6669
References 2:
Kim GY et al. The Korean Journal of Pathology. 2002; 36:5154
References 3:
Kojima M et al. Modern Pathology. 2002; 15(7):750758
References 4:
Yokozaki H et al. Japanese Journal of Clinical Oncology. 2000; 30:101104
The CD34 antigen is a single chain transmembrane glycoprotein with a molecular weight of 110 kD. The CD34 protein is selectively expressed on human lymphoid and myeloid hemopoietic progenitor cells. The CD34 antigen is also expressed on vascular enothelium. Enzyme digestion of paraffin sections is recommended with clone QBEnd/10 in perference to heat induced epitope retrieval as it produces stronger staining and reduces background elastin staining
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
QBEnd/10
Concentration:
Greater than or equal to 261 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Tornoczky T et al. Journal of Clinical Pathology. 2003; 56(5):363367
References 2:
Cox G et al. Lung Cancer. 2000; 29(3):169177
References 3:
Dhillon AP et al. Journal of Pathology. 1990; 162:274
References 4:
Ramani P et al. Histopathology. 1990; 17(3):237242
References 5:
Watanabe T et al. Journal of Clinical Pathology. 2001; 54:631636.
Chromogranin A is a 68 kD acidic protein which is reported to be widely expressed in neural tissues and in secretory granules of human endocrine cells, for example, parathyroid gland, adrenal medulla, anterior pituitary gland, islet cells of the pancreas and C cells of the thyroid. Chromogranin A expression has been reported in neuroendocrine tumors such as pituitary adenomas, islet cell tumors, phaeochromocytomas, medullary thyroid carcinomas, Merkel cell tumors and carcinoids.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
5H7
Concentration:
Greater than or equal to 13 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Khandeparker SGS et al. Journal of Pediatric Neurosciences. 2013; 8(3): 239-242.
References 2:
Marcu M et al. Current Health Sciences Journal. 2010; 32: 37-42
CA125 antigen is usually associated with ovarian epithelial malignancies. Serum assays are widely used to detect this protein in the monitoring of ovarian cancers. CA125 antigen may also be detected by immunohistochemistry and expression has been found in neoplasms such as seminal vesicle carcinoma and anaplastic lymphoma. CA125 antigen is not found exclusively in malignant tumors. CA125 is also known as MUC16.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
Ov185:1
Concentration:
Greater than or equal to 12.5 mg/L
Storage buffer:
Purified in PBS, 1% BSA and 15 mM sodium azide
Storage:
2-8°C
References 1:
Weng D et al. Int. Journal of Cancer. 2011; 129:1990-2001
References 2:
Gabriel M et al. Gynakol Geburtshilfliche Rundsch. 2000; 40(3-4):140-144
References 3:
Gabriel M et al. Ginekol Pol. 1999; 70(11):819-823
Clone C241:5:1:4 reacts specifically with Sialyl Lewisa - containing glycolipids, showing no crossreaction with Lewisa, Lewisb, or other structurally related molecules. The epitope recognized by NCL-L-CA19-9 is designated CA19-9
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
C241:5:1:4
Concentration:
n/a
Storage buffer:
Purified in PBS, 1% BSA and 15 mM sodium azide
Storage:
2-8°C
References 1:
Johansson C et al. Tumour Biology. 12: 159170 (1991)
References 2:
Kuusela P et al. British Journal of Cancer. 63: 636640 (1991)
References 3:
Haglund C et al. British Journal of Cancer. 60: 845851 (1989)
Alpha Fetoprotein (AFP) is an oncofetal antigen of 70 kD found in body fluids, which if detected in high concentrations has clinical implications. AFP is expressed in fetal liver but is not present under normal circumstances in healthy adult tissues. It is reported to be expressed in a proportion of germ cell tumors, with high frequency in yolk sac tumors.Human alpha fetoprotein. Also reacts with pig and dog alpha fetoprotein. Does not react with mouse, rat, cat or cow alpha fetoprotein
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
C3
Concentration:
Greater than or equal to 41 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Schmelzer E et al. Biotechnology and Bioengineering. 2009; 103(4): 817-827
References 2:
Piper Hanley K et al. The Journal of Biological Chemistry. 2008; 283(20): 14063-14071
References 3:
Sentani K et al. Modern Pathology. 2008; 21: 464-475
References 4:
Kielman MF et al.Nature Genetics. 2002; 32: 594-605
The CD163 molecule is a type I membrane protein also known as M130 antigen, Ber-Mac3, Ki-M8 or SM4. CD163 protein is restricted in its expression to the monocytic/macrophage lineage. It is reported to be present on all circulating monocytes and most tissue macrophages except those found in the mantle zone and germinal centers of lymphoid follicles, interdigitating reticulum cells and Langerhans cells. In addition, multi-nucleated cells within inflammatory lesions are reported not to express CD163 protein. The protein is upregulated by glucocorticoids and downregulated by the immunosuppressant cyclosporin A and by phorbol esters, while lipopolysaccharide, an inflammatory mediator, has no influence on expression. It has been proposed that a specific release mechanism of soluble CD163 antigen by human monocytes may play an important role in modulating inflammatory processes.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
10D6
Concentration:
Greater than or equal to 49 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Bronkhorst IH et al. Investigative Ophthalmology and Visual Science. 2011; 52(2):643-650
References 2:
Lau SK et al. American Journal of Clinical Pathology. 2004; 122(5):794-801
The CD68 molecule is a 110 kD intracellular glycoprotein primarily reported to be associated with cytoplasmic granules and to a lesser extent the membranes of macrophages. Markers to CD68 antigen are the most frequently used for the identification of macrophages in immunohistochemistry; however, CD68 is also found in monocytes, neutrophils, basophils and large lymphocytes. The function of the CD68 molecule is not certain but these lysosomal membrane proteins are major components and may protect the membranes from attack by acid hydrolases. It is unclear if the surface-associated CD68 protein is functionally significant or due to leakage from the lysosomes. CD68 protein expression has been demonstrated in stimulated T cells and NK cells and non-hematopoietic tissues such as liver and renal tubules.
Antibody Isotype:
IgG2a, kappa
Monosan Range:
MONXtra
Clone:
514H12
Concentration:
Greater than or equal to 37 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Gu M et al. Annals of Diagnostic Pathology. 2007; 11:64-67
References 2:
Da Costa CET et al. The Journal of Experimental Medicine. 2005; 201(5):687-693
The CD45 antigen (leukocyte common antigen) is a family of five or more high molecular weight glycoproteins present on the surface of the majority of the human leukocytes (including lymphocytes, monocytes and eosinophils) but absent from erythrocytes and platelets. Various isoforms of CD45 are generated by alternative splicing of three exons. Expression of CD45 is necessary for signaling through the T cell receptor.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
X16/99
Concentration:
Greater than or equal to 64 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Sylvester KG et al. Wound Repair and Regeneration. 2000; 8(1):36-44
References 2:
Kauma SW et al. The Journal of Clinical Endocrinology and Metabolism. 1999; 84(6):2188-2194
References 3:
Oliveira E et al. Revista da FML. 1999; 4(Supl.3):29-34
The CD43 antigen is expressed on the membrane and in the cytoplasm of T cells and cells of myeloid lineage. The molecule itself exhibits molecular weight heterogeneity with bands of 90 to 140 kD observed on SDS-PAGE between different cell lines. Cells expressing the CD43 antigen are reported to include normal and neoplastic T cells. A small proportion of B cell chronic leukemias and diffuse large B cell lymphomas are also reported to express CD43 antigen. Enzyme pretreatment may enhance staining with clone MT1 in some cases.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
MT1
Concentration:
Greater than or equal to 21 mg/L
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Higgins RA et al. Archives of Pathology and Laboratory Medicine 2008; 132:441-461
References 2:
Goteri G et al. Journal of Oral Pathology Medicine 2006; 35:254-256
The CD38 molecule is a type II single transmembrane glycoprotein with a molecular weight of 46 kD. It is an ectoenzyme with the activities of ADP-ribosyl cyclase, cyclic ADP-ribose hydrolase, NAD glycohydrolase and is involved in both the formation and hydrolysis of cADPR, a second messenger that regulates the mobilization of intracellular Ca2+ ions. Although the CD38 molecule was originally identified as a T lymphocyte differentiation antigen, it is reported to be expressed in a wide range of cells and tissues. CD38 antigen can deliver potent growth and differentiation signals to lymphoid and myeloid cells. It is found on immature cells of the B and T cell lineages but not on most mature resting peripheral lymphocytes. It is also present on thymocytes, pre-B cells, germinal center B cells, mitogen-activated T cells, Ig-secreting plasma cells, monocytes, NK cells, erythroid and myeloid progenitors in the bone marrow and brain cells. CD38 antigen has also been reported in neurofibrillary tangles, the pathological indicator of Alzheimer's disease that occurs in the neuronal perikarya and proximal dendrites.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
SPC32
Concentration:
Greater than or equal to 46 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Krukemeyer MG et al. Transplantationsmedizin. 2003; 15:4046
The CD23 molecule is the low affinity IgE receptor found on B cells.It is a membrane glycoprotein of 45 kD and is reported to be found on a sub-population of peripheral blood cells, B lymphocytes and on EBV-transformed B lymphoblastoid cell lines.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
1B12
Concentration:
Greater than or equal to 67 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Linderoth J et al. Clinical Cancer Research 2003; 9:722-728
References 2:
Peh SC et al. Singapore Medical Journal 2003; 44(4):185-191
References 3:
Maeda K et al. The Journal of Histochem. and Cytochem.2002; 50(11):1475-1485
References 4:
Watson P et al. Histopathology 2000 36(2), 145-150
CD21 antigen is a type I integral membrane glycoprotein of molecular weight 140 kD, which functions as the receptor for the C3d fragment of the third complement component. The CD21 molecule, present on mature B cells, is involved in transmitting growth-promoting signals to the interior of the B cell and acts as a receptor for Epstein-Barr virus. CD21 antigen is reported to be found in B cell chronic lymphocytic leukemias and in a subset of T cell acute lymphocytic leukemias but is absent on T lymphocytes, monocytes and granulocytes. CD21 antigen is also reported to be expressed in follicular dendritic cells and in follicular and mantle cell lymphomas, mature leukemias and lymphomas.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
2G9
Concentration:
Greater than or equal to 326 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Dupin N et al. Proceedings of the National Academy of Sciences USA. 1999; 96(8); 45464551.
The CD20 antigen is a non-glycosylated phosphoprotein of approximately 33kD which is expressed on normal and malignant human B cells and is thought to act as a receptor during B cell activation and differentiation. CD20 antigen has been reported to be expressed on normal B cells from peripheral blood, lymph node, spleen, tonsil, bone marrow, acute leukemias and chronic lymphocytic leukemias.An intracytoplasmic epitope localised on the human CD20 molecule. Reacts predominantly with a 33 kD polypeptide, but also with a minor component of 30 kD.
Antibody Isotype:
IgG2a, kappa
Monosan Range:
MONXtra
Clone:
L26
Concentration:
Greater than or equal to 95 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Mason DY et al. American Journal of Pathology. 1990; 136(6):12151222
References 2:
Cartun RW et al. American Journal of Pathology. 1987; 129(3):415421
References 3:
Norton AJ and Isaacson PG. Journal of Clinical Pathology. 1987; 40:14051412
References 4:
Ishii Y et al. Clinical Experimental Immunology. 1984; 58:183192
Prokaryotic recombinant protein corresponding to the external domain of the CD16 molecule, common to both the transmembrane form and the GPI-linked form.
CD16 antigen has a molecular weight of 50 to 70 kD and is a low affinity Fc receptor for complexed IgG, Fc/gamma RIII, expressed on natural killer (NK) cells, granulocytes, activated macrophages and a subset of T cells expressing alpha-beta or gamma-delta T cell antigen receptors. The CD16 antigen exists both as a glycosyl-phosphatidylinositol (GPI)-anchored protein in polymorphonuclear cells and as a transmembrane protein in NK cells.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
2H7
Concentration:
Greater than or equal to 18 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Qubaja M et al. Virchows Archiv. 2009; 454(4):411-419
References 2:
Wicherek L et al. Reproductive Biology and Endocrinology. 2006; 14; 4:41
References 3:
Lee SF et al. Molecular and Cell Biology. 1999; 19(11):7399-7409
CD13 antigen, also known as aminopeptidase N, is a member of type II integral membrane metalloproteases, which also includes the leukocyte antigens CD10, CD26, CD73 and BP-1. CD13 antigen is a receptor for the coronaviruses which cause respiratory disease in humans and several animal species. The antigen functions as a zinc-binding metalloprotease which plays a role in cell surface antigen presentation by trimming the N-terminal amino acids from MHC class II-bound peptides. CD13 antigen is reported to be expressed on granulocytes, monocytes and their precursors, most acute myeloid leukemias and a smaller proportion of acute lymphoid leukemias. Non-hematopoietic cells which express CD13 antigen include epithelial cells, renal proximal tubules, intestinal brush border, endothelial cells, fibroblasts, brain cells, bone marrow, osteoclasts and cells lining the bile canaliculi.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
38C12
Concentration:
Greater than or equal to 19 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Terauchi M et al.BMC Cancer 2007; 7:140
References 2:
Agis H et al. Journal of Clinical Pathology 2006; 59 (4):396-402
References 3:
Röcken C et al. Journal of Clinical Pathology 2005; 58 (10):1069-1075
CD10 antigen, also called neprilysin, is a 100 kD cell surface metalloendopeptidase which inactivates a variety of biologically active peptides. It was initially identified as the common acute lymphoblastic leukemia antigen (CALLA) and was thought to be tumor-specific. Subsequent studies, however, have shown that CD10 antigen is expressed on the surface of a wide variety of normal and neoplastic cells. In other lymphoid malignancies, CD10 antigen is reported to be expressed on cells of lymphoblastic, Burkitt's and follicular lymphomas. CD10 antigen has been identified on the surface of normal early lymphoid progenitor cells, immature B cells within adult bone marrow and germinal center B cells within lymphoid tissue. It is also expressed in various non-lymphoid cells and tissues, such as breast myoepithelial cells, bile canaliculi, fibroblasts, with especially high expression on the brush border of kidney and gut epithelial cells. (G. McIntosh et al. American Journal of Pathology. 154(1): 77-82 (1999)).
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
56C6
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Millar EK et al. Journal of Clinical Pathology 1999 52, 849-850
References 2:
McIntosh GG et al. American Journal of Pathology 1999 154(1), 7782
References 3:
Kaufmann O et al. American Journal of Clinical Pathology 1999 111(1), 117-122
References 4:
Endoh Y et al. Human Pathology 1999 30(7), 826-832
References 5:
Chu P and Arber DA. American Journal of Clinical Pathology 2000 113(3), 374382
The CD8 molecule is composed of two chains and has a molecular weight of 32 kD. It is found on a T cell subset of normal cytotoxic/suppressor cells which make up approximately 20-35% of human peripheral blood lymphocytes.The CD8 antigen is reported to be detected on natural killer cells, 80% of thymocytes, on a subpopulation of 30% of peripheral blood null cells and 15-30% of bone marrow cells.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
4B11
Concentration:
Greater than or equal to 28 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Kemp RA et al. Journal of Experimental & Clinical Cancer Research. 2011;30(1):78
References 2:
Michel S et al. British Journal of Cancer. 2008;99(11):1867-1873
References 3:
Williamson SLH et al. American Journal of Pathology 1998,152(6):1421-1426
CD5 antigen is reported to be expressed on 95% of thymocytes and 72% of peripheral blood lymphocytes. In lymph nodes, the main reactivity is observed on T cells. CD5 antigen is also expressed by many T cell leukemias, lymphomas, activated T cells and on a subset of B cells located primarily in the mantle zones of normal lymph nodes. CD5 antigen expression is also reported in T cell acute lymphocytic leukemias (T-ALL), some B cell chronic lymphocytic leukemias (B-CLL) as well as B and T cell lymphomas.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
4C7
Concentration:
Greater than or equal to 76 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Leong FJW et al. The Journal of Histotechnology. 2002; 25(4):215-227
References 2:
Walsh R et al. Arch. of Pathol.and Lab.Medicine. 2001; 125(6):781-784
References 3:
Tateyama H et al. American Journal of Clinical Pathology. 1999; 111(2):235-240
References 4:
Dorfman DM & Shahsafaei A. Modern Pathology. 1997; 10(9):859-863
References 5:
Kaufmann O et al. American Journal of Clinical Pathology. 1997; 108(6):669-673
The CD4 molecule (T4) is a single chain transmembrane glycoprotein with a molecular weight of 59 kD. The CD4 antigen is expressed on a T cell subset (helper/inducer) representing 45% of peripheral blood lymphocytes and at a lower level on monocytes and germinal center macrophages. Most cases of cutaneous T cell lymphoma, including mycosis fungoides, express the CD4 antigen and HTLV-1 associated adult T cell leukemia/lymphoma is also generally CD4 positive.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
4B12
Concentration:
Greater than or equal to 405 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Arnould L et al. British Journal of Cancer 2006; 94:259-267
References 2:
Choi HJ et al. British Journal of Dermatology 2006; 154:419-425
References 3:
Lapperre TS et al. Thorax 2006; 61:115-121
References 4:
Willemse BWM et al. Respiratory Research 2005; 6:38
References 5:
Suzuki A et al. Journal of Pathology 2002; 196:37-43
Clone MTB1 detects cortical thymocytes, Langerhans cells in epidermis, interdigitating cells of dermis and interdigitating cells of stratified squamous epithelium of tonsil. Clone MTB1 may also detect small focal groups of lymphocytes outside the germinal centers of tonsil indicating a cross-reaction with CD1b antigen.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
MTB1
Concentration:
Greater than or equal to 16 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Dultra FK et al. Journal of Oral Pathology and Medicine. 2012; 41(1):47-53
References 2:
Natamoto Y et al. Clinical and Experimental Immunology. 2007; 147:296-305
References 3:
Hubert P et al. Journal of Pathology. 2005; 206:346-355
References 4:
Rho NK et al. British Journal of Dermatology. 2004; 151:119-125
References 5:
Soilleux EJ et al. Journal of Pathology. 2001; 195(5):586-592
The Ki67 antigen is a nuclear protein which is expressed in all active parts of the cell cycle (G1, S, G2 and mitosis) but is absent in resting cells (G0). In contrast to many other cell cycle-associated proteins, the Ki67 antigen is consistently absent in quiescent cells and is not detectable during DNA repair processes. Thus, the presence of Ki67 antigen is strictly associated with the cell cycle and confined to the nucleus, suggesting an important role in the maintenance and/or regulation of the cell division cycle.
The D-type cyclins are a family of proteins which function primarily by regulating the activity of cyclin dependent kinases in the G1 phase of the cell cycle. Cyclin D1, a protein of 36 kD, is also known as PRAD1 or bcl-1. Maximum expression of cyclin D1 occurs at a critical point in mid to late G1 phase of the cell cycle. The cyclin D1 gene, located on 11q13 has been reported to be overexpressed in mantle cell lymphomas due to the chromosomal translocation t(11;18).
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
P2D11F11
Concentration:
Greater than or equal to 19 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
McIntosh GG et al. Oncogene. 1995; 11:885891
References 2:
Saiz AD et al. Journal of Pathology. 2002; 198(2):157162
References 3:
Mommers ECM et al. Journal of Pathology. 2001; 194(3):327333
References 4:
Saito T et al. Journal of Pathology. 2001; 195(2):222228
References 5:
Sheyn I et al. Human Pathology. 1997; 28(3):270276
The c-kit proto-oncogene encodes a transmembrane receptor with tyrosine kinase activity, c-kit (CD117), which is closely-related to the platelet-derived growth factor receptor family. c-kit plays a role during hematopoiesis, gametogenesis and melanogenesis. The expression of CD117 antigen is of particular interest in the study of gastrointestinal stromal tumors (GIST), small lung cell carcinomas and in melanomas.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
T595
Concentration:
Greater than or equal to 30 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Sawyer EJ et al. Journal of Pathology. 2003; 200:5964
Bcl-2 is a member of a family of proteins that are involved in apoptosis. Bcl-2 is an integral inner mitochondrial membrane protein of 25 kD and has a wide tissue distribution. It is considered to act as an inhibitor of apoptosis. For this reason, bcl-2 expression is inhibited in germinal centers where apoptosis forms part of the B cell production pathway.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
bcl-2/100/D5
Concentration:
Greater than or equal to 56 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Von Haefen C et al. Oncogene. 2002; 21(25):4009-4019
References 2:
Takes RP et al.Journal of Pathology. 2001; 194:298-302
References 3:
Tweddle DA et al. American Journal of Pathology. 2001; 158(6): 2067-2077
References 4:
Ramani P et al. Journal of Pathology. 1994; 172:273-278
References 5:
Pezzella F et al. American Journal of Pathology. 1990; 137(2):225-232
The gene encoding WAF1, also termed p21, is transcriptionally regulated by the suppressor protein, p53. Overexpression of WAF1 is growth suppressive, possibly by inhibiting the activity of cyclin/CDK complexes. One consequence of WAF1 binding to cyclin/CDK is the inhibition of Rb protein phosphorylation. Induction of WAF1 expression requires wild type p53 activity in cells undergoing p53 dependent G1 arrest or apoptosis. Mutation of the p53 gene is a common event in human cancer and results in the failure to produce WAF1. The effect of this may lead to uncontrolled cell proliferation.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
4D10
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15 mM sodium azide
Storage:
2-8°C
References 1:
Göhring UJ et al. Journal of Clinical Pathology. 54: 866870 (2001)
References 2:
Schwerer MJ et al. Journal of Clinical Pathology. 54: 871876 (2001)
References 3:
Tweddle DA et al. American Journal of Pathology. 158 (6): 20672077 (2001)
References 4:
Garcia JF et al. Histopathology. 30: 120125 (1997)
Retinoblastoma (Rb) is a rare tumor of the retina associated with mutations of chromosome 13. The nuclear phosphoprotein encoded by the Rb tumor suppressor gene is present in many cells and may indirectly regulate cell growth by activating the transcription factor ATF-2. Activation of ATF-2 initiates expression of TGF-beta2, which in turn inhibits transcription of genes affecting cell growth. Bilateral mutation of the Rb gene may potentially play a role in the development of a number of malignant tumors.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
13A10
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with 15mM sodium azide
Storage:
2-8°C
References 1:
Jares P et al. Journal of Pathology. 182: 160-166 (1997)
References 2:
Karpeh MS et al. British Journal of Cancer. 72: 986-991 (1995)
References 3:
Stefanini M et al. Nature. 216: 173-174 (1967)
References 4:
Bartek J et al. Oncogene. 7: 101-108 (1992)
References 5:
Sanders BM et al. British Journal of Cancer. 60: 358-365 (1989)
This antibody reacts with a subset of B-lymphocytes localized in the follicular mantle zone. It reacts with 97% of hairy cell leukemia cases. This antibody shows strong positive staining of about 35% of cases of high grade B cell lymphomas. Positive control Tonsil.
GATA binding protein 3 or GATA3, is a zinc finger transcription factor and plays an important role in promoting and directing cell proliferation, development, and differentiation in many tissues and cell types.1 The human GATA3 gene has been mapped to chromosome 10p15.3 GATA3 expression is primarily seen in breast carcinoma and urothelial carcinoma. Anti-GATA3 can also be useful in the identification of unknown primary carcinoma when carcinomas of the breast or bladder are a possibility
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
L50-823
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Higgins JP, et al. Am J Surg Pathol. 2007; 31:673-80
DOG1 is expressed ubiquitously in gastrointestinal stromal tumors irrespective of c-kit or PDGFR alpha mutation status. Reactivity for DOG1 may aid in the diagnosis of gastrointestinal stromal tumors, including PDGFRA mutants that fail to express c-kit antigen, and lead to appropriate treatment with imatinib mesylate, an inhibitor of the c-kit tyrosine kinase. Positive control Gastrointestinal stromal tumor tissue
CD4 is a 55 kD glycoprotein expressed on the surface of T-helper/regulatory T-cells, monocytes, macrophages, and dendritic cells. Anti-CD4 is used in the immunophenotyping of lymphoproliferative disorders.1 The majority of peripheral T-cell lymphomas are derived from the T-helper/regulatory cell subset so that most mature T-cell neoplasms are CD4+ CD8-.2 As with other T-cell antigens, CD4 may be aberrantly expressed in neoplastic T-cells so that the evaluation of such tumors requires the application of a panel of markers in order to identify tumors with CD4 aberrant expression.1-3
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP204
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Leong AS-Y, et al. Greenwich Medical Media Ltd. 2003
References 2:
Akiyama T, et al. Pathol Int. 2008; 58:626-34
References 3:
Garcia-Herrera A, et al. J Clin Oncol. 2008; 26:3364-71
Carbonic Anhydrase (CA IX) is part of a family of zinc containing metalloproteins that catalyze the reversible hydration of CO2.1 Among these, CA IX is anchored to the cell membrane and is expressed in the human gastrointestinal tract, chiefly in the stomach.1 Data suggest consistent immunoreactivity for anti-CA IX in clear cell renal cell carcinoma (RCC) making it a useful marker for this type of tumor.2
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP161
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Leppilampi M, et al. World J Gastroenterol. 2003; 9:1398-1403
Adenovirus infection is associated with a broad spectrum of clinical disease in both children and adults. It has gained more attention as an important complication in patients who have undergone bone marrow or solid organ transplantation. The incidence of adenovirus infection in bone marrow transplant patients has been reported at 5-20%. Adenovirus infection on morphology should be differentially diagnosed from other virus infections, especially CMV infection. Anti-adenovirus can assist in this differential diagnosis by showing a round or crescent-shaped nuclear inclusion, generally within the surface epithelium and is exclusively intra-nuclear in location.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
20/11 & 2/6
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
son, MG. Clin Infect Dis. 2006; 43: 3319
References 2:
Shayan K, et al. Arch Pathol Lab Med 2003;127:1615-8
PAX-8 is a transcription factor expressed during embryonic development of Müllerian organs, kidney, and thyroid, with continued expression in some epithelial cell types of these mature tissues.1 It can be useful for marking several types of carcinoma including ovarian serous carcinoma, clear cell renal cell carcinoma, and papillary thyroid carcinoma.1-5 Additionally, PAX-8 is not found in the epithelial cells of the breast, lung, mesothelium, stomach, colon, pancreas and other sites.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-50
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ozcan A, et al. Mod Pathol. 2011; 24:751-64
References 2:
Laury AR, et al. Am J Surg Pathol. 2011; 35:816-26
References 3:
Nonaka, D et al. Am J Surg Pathol. 2008; 32:1566-71
Anti-Cytokeratin 5 & 6 is a marker for eptihelioid mesotheliomas. Anti-Cytokeratin 5 & 6 stains the cytoplasm of such cells. Anti-TTF-1 stains the nuclei in the case of lung adenocarcinomas and is negative in nearly all mesotheliomas. The nuclear vs. cytoplasmic staining pattern of this cocktail can be useful in distinguishing between mesothelioma and adenocarcinoma of the lung.
Antibody Isotype:
IgG1/IgG1/IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
D5/16B4 & 8G7G3/1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ordóñez NG. Am J Surg Pathol. 1998; 22:1215-21
References 2:
Jang KY, et al. Anal Quant Cytol Histol; 2001; 23:400-4
References 3:
Srodon M, et al. Hum Pathol. 2002; 33:642-5
References 4:
Abutaily AS, et al. J Clin Pathol. 2002; 55:662-8
References 5:
Bejarano PA, et al. Arch Pathol Lab Med. 2003; 127:193-5
SOX-11 which is a member of the SOX (SRY-related HMG-box) family is a transcription factor normally expressed in the developing human central nervous system and plays a role in embryonic cell determination. Studies show that SOX-11 can be used as a marker for mantle cell lymphoma (MCL). Mantle cell lymphoma (MCL) accounts for 5% to 10% of mature B-cell neoplasms and is an aggressive disease genetically characterized by overexpression of cyclin D1 (CCND1), an important regulator of the G1/S phase of the cell cycle, due to the specific translocation t(11;14)(q13;q32).
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-58
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Claudins are a family of over twenty proteins which are components of tight junctions. Tight junctions are specialized regions of cell-to-cell contact made up of a network of strands to act as a molecular gasket for preventing the leakage of ions, water, etc., between cells.1 Claudin 1 has been shown to distinguish epithelial neoplasms from lymphomas, making it a useful marker for nearly all carcinomas.2
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Folpe AL, et al. Am J Surg Pathol. 2002; 26:1620-6
The antibody abels a 50kDa, multiple myeloma oncogen-1 (MUM1) protein. MUM1 is encoded by the MUM1/IRF-4 gene, which is mapped to 6q23-25 and identified as a myeloma-associated oncogene. It is a member of the interferon regulatory factor family of transcription factors and plays an important role in the regulation of gene expression in response to signaling by interferon and other cytokines. MUM1 positive cells express the protein in the nucleus in a diffuse and microgranular pattern. However, some positivity is also observed in the cytoplasm of MUM1-expressing cells. In normal/reactive lymphoid tissues, such as lymph node, this antibody stains plasma cells, some B-cells in the light zone of terminal centers, and a subset of T-cells (T-cells in germinal centers and interfollicular areas). Anti-MUM1 antibody can stain other B-cell lymphomas such as lymphoplasmacytic lymphoma, chronic lymphocytic leukemia, follicular lymphoma, marginal zone lymphoma, lymphoblastic lymphoma /leukemia, primary effusion lymphoma, DLBCL, Burkitt-like lymphoma, and classical Hodgkin lymphoma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-43
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Grossman A, et al. Genomics. 1996; 37:229-33
References 2:
Neresh KN. Haematologica. 2007; 92:267-8
References 3:
Van Imhoff GW, et al. J Clin Oncol. 2006; 24:4135-42
References 4:
Gualco G, et al. Appl Immunohistochem Mol Morphol. 2010; 18:301-10
Glucose transporter type I (GLUT1), a prototype member of GLUT super family, reacts with a 55 kD protein, is a membrane-associated erythrocyte glucose transport protein. It is a major glucose transporter in the mammalian blood-brain barrier, and also mediates glucose transport in endothelial cells of the vasculature, adipose tissue and cardiac muscle. GLUT1 is detectable in many human tissues including those of colon, lung, stomach, esophagus, and breast. GLUT1 is overexpressed in malignant cells and in a variety of tumors that include the breast, pancreas, cervix, endometrium, lung, mesothelium, colon, bladder, thyroid, bone, soft tissues, and oral cavity. Immuohistochemical detection of GLUT1 can discriminate between reactive mesothelium and malignant mesothelioma. Anti-GLUT1 with anti-Claudin1, and anti-EMA are perineurial markers in diagnosis of perineuriomas. Anti-GLUT1 is also useful in distinguishing benign endometrial hyperplasia from atypical endometrial hyperplasia and adenocarcinoma. GLUT1 expression has been associated with increased malignant potential, invasiveness, and a poor prognosis in general. Expression of GLUT1 is a late event in colorectal cancer and expression in a high proportion of cancer cells is associated with a high incidence of lymph node metastases.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kato Y, et al. Mod Pathol. 2006; 20:215-20
References 2:
Afify A, et al. Acta Cytol. 2005; 49:621-6
References 3:
Parente P, et al. J Exp Clin Cancer Res. 2008; 27:34
Transcription factor E3 (TFE3) is a protein expressed in many cell types that are encoded by the TFE3 gene. This gene may be involved in chromosomal translocations that occur in some cancers.Xp11 translocation renal cell carcinomas (RCC) are a recently recognized subset of RCC, characterized by chromosome translocations involving the Xp11.2 break point and resulting in gene fusions involving the TFE3 transcription factor gene that maps to this locus.1 Alveolar soft part sarcoma (ASPS) is an uncommon soft tissue sarcoma of uncertain differentiation. The hallmark of ASPS is a chromosomal rearrangement at 17q25 and Xp11.2 engendering an ASPSCR1-TFE3 fusion gene responsible for an aberrant transcription factor presumably enabling pathogenesis.1-5
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-37
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Argani P. Am J Clin Pathol. 2006; 126:332-4
References 2:
Argani P, et al. Am J SurgPathol. 2003; 27:750-61
References 3:
Argani P, et al. Clin Lab Med.2005; 25:363-78
References 4:
Lazar AJ, et al. Histopathol. 2009; 55:750-5
References 5:
Lin G, et al. Arch Pathol Lab Med. 2015; 139:106-21
Napsin is a pepsin-like aspartic proteinase in the A1 clan of the AA clade of proteinases. There are two closely related napsins, napsin A (NAPSA) and napsin B (NAPSB). Napsin A is involved in processing propeptide pulmonary surfactant protein B (proSP-B) in the lung. In normal tissue, Napsin A is expressed in type II pneumocytes of the lung and proximal tubules of the kidney. Napsin A is a useful marker for lung adenocarcinoma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-60
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Jagirdar J. Arch Pathol Lab Med.; 132:384-96 (2008)
References 2:
Bishop JA, et al.. Hum Pathol.; 41:20-5 (2010)
References 3:
Ye J, et al. Appl Immunohistochem Mol Morphol.; 19:313-17 (2011)
References 4:
Mukhopadhyay S, et al. Am J Surg Pathol.; 35:15-25 (2011)
References 5:
Rawlings ND and Salvesen GS. Academic Press.; p.69-71 (2013)
Napsin is a pepsin-like aspartic proteinase in the A1 clan of the AA clade of proteinases. There are two closely related napsins, napsin A (NAPSA) and napsin B (NAPSB). Napsin A is involved in processing propeptide pulmonary surfactant protein B (proSP-B) in the lung.4 In normal tissue, Napsin A is expressed in type II pneumocytes of the lung and proximal tubules of the kidney. Napsin A is a useful marker for lung adenocarcinoma
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Brasch F, et al. J Biol Chem. 2003; 278: 49006-14
References 2:
Jagirdar J et al. Arch Pathol Lab Med. 2008; 132:384-96
References 3:
Bishop JA, et al. Hum Pathol. 2010; 41:20-5
References 4:
Ye J, et al. Appl Immunohistochem Mol Morphol. 2011; 19:313-17
References 5:
Mukhopadhyay S, et al. Am J Surg Pathol. 2011; 35:15-25
Thrombomodulin is a transmembrane glycoprotein composed of 575 amino acids (molecular weight 75 kD) with natural anticoagulant properties. It is normally expressed by a restricted number of cells, such as endothelial and mesothelial cells. In addition, synovial lining and syncytiotrophoblasts of human placenta also express thrombomodulin. Antithrombomodulin has demonstrated positivity in benign vascular tumors such as hemangioma and most malignant vascular tumors (Kaposis sarcoma and epitheliod hemangioendothelioma). Hence, anti-thrombomodulin serves as a sensitive marker for lymphatic endothelial cells and their tumors. There has also been recent interest in the use of antithrombomodulin as an immunohiostochemical marker for mesothelial cells and malignant mesotheliomas. Anti-thrombomodulin is immunoexpressed in a variety of other tumors including urothelial cell carcinomas
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
1009
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Acebo E, et al. Histol. Histopath. 2001; 16:1031-6
References 2:
Appleton MA, et al. Histopathology. 1996; 29:153-7
References 3:
Attanoos RL, et al. Histopathology. 1996; 29:209-15
References 4:
Attanoos RL, et al. Histopathology. 2001; 39:584-8
The antibody reacts with neuroendocrine cells of human adrenal medulla, carotid body, skin, pituitary, thyroid, lung, pancreas, and gastrointestinal mucosa. This antibody identifies normal neuroendocrine cells and neuroendocrine neoplasms. Diffuse, finely granular, cytoplasmic staining is observed, which probably correlates with the distribution of the antigen within neurosecretory vesicles. The expression of synaptophysin is independent of the presence of NSE or other neuroendocrine markers. Antisynaptophysin is an independent, broad-range marker of neural and neuroendocrine differentiation.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-40
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Wiedenmann, B, et al. Cell 1985;41:1017-1028
References 2:
Navone, F et al. J Cell Biol 1986;103:2511-2527
References 3:
Lyda MH et al. Hum Pathol. 2000 Aug;31(8):980-7
References 4:
Skacel M et al. Appl Immunohistochem Mol Morphol. 2000 Sep;8(3):302-9
The TRIS/EDTA buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 9.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 9.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of TRIS?EDTA buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the TRIS/EDTA buffer and slides to 98°C and incubate for 15 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
The TRIS/EDTA buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 9.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Principle of method:
Heat-Induced Epitope Retrieval
Reagents provided:
100 ml 10x concentrate solution. TRIS/EDTA buffer, Heat-Induced Epitope Retrieval (10x, Tween20), Blue
Storage and handling:
Store at room temperature
Reagent preparation:
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 9.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of TRIS?EDTA buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the TRIS/EDTA buffer and slides to 98°C and incubate for 15 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
The Citrate buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 6.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Principle of method:
Heat-Induced Epitope Retrieval
Reagents provided:
100 ml 10x concentrate solution. Citrate buffer, Heat-Induced Epitope Retrieval (10x, Tween20), Red
Storage and handling:
Store at room temperature
Reagent preparation:
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 6.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of citrate buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the citrate buffer and slides to 98°C and incubate for 20 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
The Citrate buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 6.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 6.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of citrate buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the citrate buffer and slides to 98°C and incubate for 20 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDAB substrate DAB (3,3Diaminobenzidine) is a widely used chromogen for immunohistochemical staining and immunoblotting. When in the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol and xylene. This product comes in a two-component system consisting of a liquid stable DAB chromogen and DAB substrate buffer. DAB Solution A: Readyto-Use Buffered H2O2,; DAB Solution B: Concentrated DAB solution. BrightDAB is a very stable and superior formulation of DAB. In some cases, antibodies titers may increase by two-fold. BrightDAB can be used both manually and on automated stainers. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests. Also available in 500 ml and 1000 ml.
Principle of method:
DAB substrate for use with HRP-labeled detection systems
Reagents provided:
DAB Solution A: Buffered H2O2 (Ready-to-Use) 110 ml DAB Solution B: Concentrated DAB solution 5 ml
Storage and handling:
2-8°C and in the dark
Reagent preparation:
1 Working solution: Add 40 ?l DAB Solution B (± one drop) to 1 ml Solution A, mix well. 2 Incubate the DAB solution one time 8 minutes without washing in between.
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, polymer HRP, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; 8 min), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system Goat Anti-Rabbit IgG HRP and Goat anti-Mouse IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system, peroxidase Goat Anti-Rabbit HRP and Goat anti-Mouse AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Rabbit and Mouse bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests.
Principle of method:
BrightVision, one step detection system Goat Anti- Rabbit HRP and Mouse AP (Ready-to-Use).
Reagents provided:
BrightVision, One step detection system Goat anti-Rabbit HRP / Goat anti-Mouse AP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse and/or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9 Detection system, polymer Rabbit HRP/Mouse AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (DAB; see applicble IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Substrate (Fast Red / New Fuchsin; see applicable IFU), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain and coverslip with aqueous mounting medium.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system Goat Anti-Mouse IgG HRP and Goat anti-rabbit IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system, peroxidase Goat Anti-Mouse HRP/Goat anti-Rabbit AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Rabbit and Mouse bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests.
Principle of method:
BrightVision, one step detection system Goat Anti- Mouse HRP and Rabbit AP (Ready-to-Use).
Reagents provided:
BrightVision, One step detection system Goat anti-Mouse HRP / Goat anti-Rabbit AP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse and/or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9 Detection system, polymer Mouse HRP/Rabbit AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (DAB; see applicble IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Substrate (Fast Red / New Fuchsin; see applicable IFU), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain and coverslip with aqueous mounting medium.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision Ultimate one step component detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP and DAB, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision Ultimate detection system, peroxidase Goat Anti-Mouse/Rabbit HRP plus DAB, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision Ultimate detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests.
Principle of method:
BrightVision Ultimate, one step detection system, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use) plus DAB.
Reagents provided:
BrightVision Ultimate plus, one detection system, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use) plus DAB 1. one step detection, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use): 55 ml 2. DAB Solution A: Buffered H2O2 (Ready-to-Use): 83 ml 4. DAB Solution B: Concentrated DAB solution: 4 ml
Storage and handling:
2-8°C and in the dark
Reagent preparation:
Preparation DAB: Add 40?l DAB Solution B (one drop) to 1 ml substrate Solution A, mix well. Volume and the quality of the Bright DAB has been formulized so they also can be used in automatic stainers, when a higher volume is required.
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9 Detection system, polymer Mouse/Rabbit HRP, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; 8 min.; Preparation DAB: Add 40?l DAB Solution B (one drop) to 1 ml substrate Solution A, mix well. Volume and the quality of the Bright DAB has been formulized so they also can be used in automatic stainers, when a higher volume is required.), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dyhydrate and coverslip (Auxiliary.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision Ultimate plus two steps component detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP and DAB, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision Ultimate plus detection system, peroxidase Goat Anti-Mouse/Rabbit HRP plus DAB, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision Ultimate plus detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests.
Principle of method:
BrightVision Ultimate plus, two steps detection system, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use) plus DAB.
Reagents provided:
BrightVision Ultimate plus, two steps detection system, Goat Anti-Mouse/Rabbit HRP (Ready-to-Use) plus DAB 1. Post-blocking (ready-to-use): 55 ml 2. Polymer Goat Anti-Mouse/Rabbit HRP (Ready-to-Use): 55 ml 3. DAB Solution A: Buffered H2O2 (Ready-to-Use): 83 ml 4. DAB Solution B: Concentrated DAB solution: 4 ml
Storage and handling:
2-8°C and in the dark
Reagent preparation:
Preparation DAB: Add 40?l DAB Solution B (one drop) to 1 ml substrate Solution A, mix well. Volume and the quality of the Bright DAB has been formulized so they also can be used in automatic stainers, when a higher volume is required.
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, post-blocking (Post-blocking; 15 min), 10 Wash buffer (PBS or TBS buffer; 2x 5 in). 11 Detection system, step 2, polymer Mouse/Rabbit HRP, (Labeled polymer; 30 min), 12. Wash buffer (PBS or TBS buffer; 2x 5 min), 13. Substrate (DAB; 8 min; Preparation DAB: Add 40?l DAB Solution B (one drop) to 1 ml substrate Solution A, mix well. Volume and the quality of the Bright DAB has been formulized so they also can be used in automatic stainers, when a higher volume is required.), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dyhydrate and coverslip (Auxiliary.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system Goat Anti-Rabbit IgG AP, is intended for use in immunohistochemistry for the detection of rabbit antibodies.
The BrightVision detection system Goat Anti-Rabbit AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided). Also available in 110 ml, 500 ml and 1000 ml.
Principle of method:
One step detection system goat anti-rabbit IgG AP
Reagents provided:
One step detection system Goat anti-Rabbit AP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary rabbit antibody (Antibody; 30 min), 8. Wash buffer (TBS buffer; 2x 5 min), 9. Detection system, polymer Rabbit AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (Fast Red / New Fuchsin; see applicable IFU), 12. Wash aqua dest (Wash; 2x 2 min), 12. Counterstain and coverslip with aqueous mounting medium.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system peroxidase Goat Anti-Rabbit IgG HRP, is intended for use in immunohistochemistry for the detection of rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti-Rabbit HRP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided). Also available in 110 ml, 500 ml and 1000 ml.
Principle of method:
One step detection system peroxidase goat anti-rabbit IgG HRP
Reagents provided:
One step detection system Goat anti-Rabbit HRP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. H2O2 (conc3%) (Tissue preparing; 10 min), 5. Wash buffer (PBS or TBS buffer; 2x 5 min), 6. Primary rabbit antibody (Antibody; 30 min), 7. Wash buffer (PBS or TBS buffer; 2x 5 min), 8. Detection system, polymer Rabbit HRP, (Labeled polymer; 30 min), 9. Wash buffer (PBS or TBS buffer; 2x 5 min), 10. Substrate (DAB; see applicable IFU), 11. Wash aqua dest (Wash; 2x 2 min), 12. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system peroxidase Goat Anti-Mouse IgG AP, is intended for use in immunohistochemistry for the detection of mouse antibodies.
The BrightVision detection system Goat Anti- Mouse AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided). Also available in 110 ml, 500 ml and 1000 ml.
Principle of method:
One step detection system peroxidase goat anti-mouse IgG AP
Reagents provided:
One step detection system Goat anti-Mouse AP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse antibody (Antibody; 30 min), 8. Wash buffer (TBS buffer; 2x 5 min), 9. Detection system, polymer Mouse AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (FAST Red / New Fuchsin; see applicable IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain and coverslip with a aqueous mounting medium (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system peroxidase Goat Anti-Mouse IgG HRP, is intended for use in immunohistochemistry for the detection of mouse antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse HRP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided). Also available in 110 ml, 500 ml and 1000 ml.
Principle of method:
One step detection system peroxidase goat anti-mouse IgG HRP
Reagents provided:
One step detection system Goat anti-Mouse HRP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, polymer Mouse, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; see applicable IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system Goat Anti-Mouse/Rabbit IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system Goat Anti- Mouse/Rabbit AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided). Also available in 110 ml, 500 ml and 1000 ml.
Principle of method:
One step detection system goat anti-mouse/rabbit IgG AP
Reagents provided:
One step detection system Goat anti-Mouse/Rabbit AP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, polymer Mouse/Rabbit AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (Fast Red / New Fuchsin, see applicable IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain and coverslip with aqueous mounting medium.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision one step detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse/Rabbit HRP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided). Also available in 110 ml, 500 ml and 1000 ml.
Principle of method:
One step detection system peroxidase goat anti-mouse/rabbit IgG HRP
Reagents provided:
One step detection system Goat anti-Mouse/Rabbit HRP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, polymer HRP, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; see applicable IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision two components detection system peroxidase Goat Anti-Mouse/Rabbit IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse/Rabbit AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided).
Principle of method:
Two steps detection system goat anti-mouse/rabbit AP
Reagents provided:
Post-blocking (ready-to-use) (Gold) 55 ml and Polymer Goat Anti-Mouse/Rabbit AP (ready-to-use) (Ruby) 55 ml.
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, post-blocking (Post-blocking; 15 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Detection system, step 2, polymer Mouse/Rabbit AP (Labeled polymer; 30 min), 12. Wash buffer (PBS or TBS buffer; 2x 5 min), 13. If applicable Substrate (DAB), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision two components detection system peroxidase Goat Anti-Mouse/Rabbit IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse/Rabbit AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided).
Principle of method:
Two steps detection system goat anti-mouse/rabbit AP
Reagents provided:
Post-blocking (ready-to-use) 55 ml and Polymer Goat Anti-Mouse/Rabbit AP (ready-to-use) 55 ml.
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, post-blocking (Post-blocking; 15 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Detection system, step 2, polymer Mouse/Rabbit AP (Labeled polymer; 30 min), 12. Wash buffer (PBS or TBS buffer; 2x 5 min), 13. If applicable Substrate (DAB), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision two components detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse/Rabbit HRP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided).
Principle of method:
Two steps detection system goat anti-mouse/rabbit HRP
Reagents provided:
Post-blocking (ready-to-use) (Gold) 55 ml and Polymer Goat Anti-Mouse/Rabbit HRP (ready-to-use) (Ruby) 55 ml.
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, post-blocking (Post-blocking; 15 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Detection system, step 2, polymer Mouse/Rabbit HRP (Labeled polymer; 30 min), 12. Wash buffer (PBS or TBS buffer; 2x 5 min), 13. If applicable Substrate (DAB), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
For in-vitro Diagnostic Use. The BrightVision two components detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse/Rabbit HRP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided).
Principle of method:
Two steps detection system goat anti-mouse/rabbit HRP
Reagents provided:
Post-blocking (ready-to-use) 55 ml and Polymer Goat Anti-Mouse/Rabbit HRP (ready-to-use) 55 ml.
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, post-blocking (Post-blocking; 15 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Detection system, step 2, polymer Mouse/Rabbit HRP (Labeled polymer; 30 min), 12. Wash buffer (PBS or TBS buffer; 2x 5 min), 13. If applicable Substrate (DAB), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
The aqua mounting medium is a widely used for coverslipping from aqueous solutions and is non-fluorescing and has an antifade component to increase the viewing time of the specimen. Use Aqua-Poly/Mount with most fluorescent dyes and stains including DAB, Alkaline Phosphatase, Fast Red, AEC (aminoethylcarbazole) and a variety of other chromogens to enhance and retain fluorescent intensity. The mounting medium (water based) can be used for frozen sections, fat stains, chromogens for immunohistochemistry and in situ hybridization as well as other applications requiring a water-soluble mounting medium.
Principle of method:
Coverslipping from aqueous solutions
Reagents provided:
50 ml Mounting medium (water based), ready-to-use.
Storage and handling:
Store at room temperature
Procedure:
1. Prepare slides as required. 2. Prior to coverslipping rinse the slides in distilled or deionized water. 3. Place the bottle upside down with the end cap attached in a container before use. This will help clear of tiny bubbles. 4. Blot excess water from the slide without drying the tissue specimen. The tissue must be moist prior to mounting. 5. Apply enough drops of aqua mounting medium on the tissue section so that the tissue is completely covered. 6. Carefully lower the coverslip at an angle while gently applying pressure to force any excess medium and air bubbles away from the tissue and out from under the coverslip. 7. Slides can be dried at room temperature or at 4°C or in an oven but the temperature hight is depending what type of staining is used. 8. Allow the aqua mounting medium to dry before examine under the microscope.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
4x 100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
Paraformaldehyde (PFA) is the polymerization product of formaldehyde with a typical degree of polymerization of 8100 units. Paraformaldehyde is not a fixative itself; it must be depolymerized to formaldehyde in solution. In cell culture, a typical formaldehyde fixing procedure would involve using a 4% formaldehyde solution in phosphate buffered saline (PBS) on ice for 10 minutes. Fixing ensures that sample cell structures stay intact and that antigens are immobilized, while ideally still permitting unfettered access of antibodies to target antigens. CAS 30525-89-4, soluble in water, pH 7.0-7.6 at 25°C. The solution should be clear, colorless, with no precipitate.
Principle of method:
Fixing cells for immunohistochemistry (IHC)
Reagents provided:
500 ml 4 % Paraformaldehyde (PFA) solution in PBS
Storage and handling:
Store at -20°C for one year
Trouble shooting:
Adjust pH to 7.2-7.6 with NaH2PO4 if pH
Precautions:
Causes serious eye damage. Suspected of causing cancer. May cause an allergic skin reaction. Causes skin irritation. Avoid breathing dust/fume/gas/mist/vapors/spray. Wear protective gloves/protective clothing/eye protection/face protection. Use personal protective equipment as required. IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. Store locked up. Dispose of contents/container in accordance with local/regional/national/international regulations
Permanent HRP Green Kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). It results in the formation of a green precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in organic solvents and can be observed by light microscopy. The HRP Green precipitate shows a very good contrast to red chromogenic substrates used with alkaline phosphatase detection systems and is therefore especially recommended for double stains. Permanent HRP Green Kit is intended for research use only.
100 ml HRP Green Substrate Buffer 3 ml HRP Green Chromogen 1 Dilution Vial
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. It is stable for at least 4 hours. Excess working solution should be disposed. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
1) Pipette 1 ml HRP Green Substrate Buffer into the provided dilution vial. 2) Add 1 drop (30 µl) of HRP Green Chromogen. Mix thoroughly. 3) The resulting working solution is stable for at least 4 hours. If you want to prepare other quantities of the working solution, please use the same ratio HRP Green Substrate and HRP Chromogen.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the Permanent HRP Green working solution onto the slide. Incubate for 2-10 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Do not exceed incubation times of 30 sec per dehydration step. Use only high purity xylene. Mount with a permanent mounting medium. Note: The colour intensity can be intensified by increasing the chromogen concentration (up to 3 drops or 90 µl chromogen) in the working solution. Lithium carbonate may have a negative effect on the staining result. We recommend to only bluing in tap water. Occasionally precipitates may appear in the HRP Green Chromogen solution. This doesnt affect the staining result.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the Permanent HRP Green chromogen, a green precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in organic solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Too long incubation steps at the final dehydration serious can diminish the staining intensity. Also low grade xylene and some forms of recycled alcohol can have a negative effect on the staining result. In double stain procedures we recommend to use Permanent HRP Green as the last chromogen. Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
AEC Single Solution is a ready-to-use solution intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous mounting media and can be observed by light microscopy. AEC Single Solution is especially useful when a high sensitivity is desired.
100 ml AEC Single Solution (ready-to-use) 2 Dropper Bottles
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. AEC Single Solution is a ready-to-use solution. Preparation of a working solution as in other chromogenic substrates is not necessary. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use. AEC Single Solution can be used directly from the refrigerator and should be stored again at 2-8°C aft er use. When using the small package (8 ml) please directly drop from the bottle. When using the 100 ml package (MON-APP209) please transfer up to 8 ml of the AEC Single Solution into one of the provided dropper bottles. The transferred solution is stable for many weeks if stored at 2-8°C. The volume required for several staining runs should be transferred so that the 100 ml stock bottle has to be opened only a few times. If you would like to pipette the solution use a clean vial from which you pipette. Remaining quantities should not be filled back into the bottle but disposed as hazardous material.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the AEC Single Solution to the slide. Incubate for 3-6 minutes. (Incubation time can be extended up to 30 minutes, if desired.) 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Mount with an aqueous mounting medium.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. The coloured precipitate formed by AEC is soluble in organic solvents. The tissue sections therefore have to be counterstained with aqueous solutions (e. g. Gills or Mayers haematoxylin) and mounted with aqueous mounting media. The colour intensity of the reaction product can decrease with time, especially when exposed to light. The staining reaction itself can be influenced in the same way when carried out in strong light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. AEC (3-Amino-9-ethylcarbazol) and the solvents used are considered hazardous materials. Material safety data sheets (MSDS) are available upon request. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Oxidising substances, e. g. metals, dust, bacteria or glass devices can influence the stability of AEC Single Solution. Such contaminations have to be avoided. Non-consumed solution needs to be discarded as dangerous substance.
The Fast AP One-Step Polymer anti-Mouse/Rabbit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It was developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE. It is intended for in vitro diagnostic use.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Fast AP One-Step Polymer anti-Mouse/Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzymesubstrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. Cross-reactivity with primary antibodies from rat has been observed. In contrast to other detection techniques, which often use the streptavidin-biotin system the Fast AP One-Step Polymer anti-Mouse/Rabbit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP OneStep Polymer is minimized by incubation with a protein blocking solution. This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the AP One-Step Polymer is applied and incubated. Any excess of unbound polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent AP Red leads to the formation of a magentared product of reaction at the place of the target antigen.
Reagents provided:
100 ml Fast AP One-Step Polymer anti-Mouse/Rabbit (ready-to-use) Substrate systems recommended: Permanaent AP Red kit Materials required but not supplied Positive and negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS Blocking Solution (for protein blocking, optional) Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagent, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Blocking Solution (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. AP One-Step Polymer anti-Mouse/Rabbit 30 Min. 6. Washing with wash buffer 3 x 2 min. 7. Permanent AP Red, 10-20 min. (Controlling the colour intensity via light microscope is recommended.) 8. Stopping the reaction with distilled H2O when the desired colour intensity is attained 9. Counterstaining and blueing 10. Mounting: permanent or aqueous with Permanent AP Red Kit
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Fast AP One-Step Polymer anti-Mouse/Rabbit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It was developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE. It is intended for in vitro diagnostic use.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Fast AP One-Step Polymer anti-Mouse/Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzymesubstrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. Cross-reactivity with primary antibodies from rat has been observed. In contrast to other detection techniques, which often use the streptavidin-biotin system the Fast AP One-Step Polymer anti-Mouse/Rabbit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP OneStep Polymer is minimized by incubation with a protein blocking solution. This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the AP One-Step Polymer is applied and incubated. Any excess of unbound polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent AP Red leads to the formation of a magentared product of reaction at the place of the target antigen.
Reagents provided:
6 ml Fast AP One-Step Polymer anti-Mouse/Rabbit (ready-to-use) Substrate systems recommended: Permanaent AP Red kit Materials required but not supplied Positive and negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS Blocking Solution (for protein blocking, optional) Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagent, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Blocking Solution (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. AP One-Step Polymer anti-Mouse/Rabbit 30 Min. 6. Washing with wash buffer 3 x 2 min. 7. Permanent AP Red, 10-20 min. (Controlling the colour intensity via light microscope is recommended.) 8. Stopping the reaction with distilled H2O when the desired colour intensity is attained 9. Counterstaining and blueing 10. Mounting: permanent or aqueous with Permanent AP Red Kit
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus AP Polymer anti-Mouse is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with monoclonal primary antibodies obtained from mouse. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Polymer anti-Mouse is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The reagent is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mouse. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus AP Polymer anti-Mouse avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP polymer is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the AP-polymer is applied and incubated. Any excess of unbound AP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Fast Red leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red), New Fuchsin (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
100 ml AP-Polymer anti-Mouse (Ready-to-use) Substrate systems recommended: Permanent AP Red kit, Fast Red substrate kit, New Fuchsin kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out
Procedure:
1. Blocking Solution (protein block, this step is optional) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. AP-polymer anti Mouse 30 min. 6. Washing with wash buffer 3 x 2 min. 7. Fast Red, Permanent AP Red, NBT/BCIP or New Fuchsin 5-15 min. (Controlling the colour intensity via light microscope is recommended.) 8. Stopping the reaction with distilled H2O when the desired colour intensity is attained 9. Counterstaining and blueing 10. Mounting: aqueous with Fast Red, permanent with Permanent AP Red, NBT/BCIP or New Fuchsin
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use a Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light.Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus AP Polymer anti-Mouse is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with monoclonal primary antibodies obtained from mouse. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Polymer anti-Mouse is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The reagent is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mouse. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus AP Polymer anti-Mouse avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP polymer is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the AP-polymer is applied and incubated. Any excess of unbound AP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Fast Red leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red), New Fuchsin (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
6 ml AP-Polymer anti-Mouse (Ready-to-use) Substrate systems recommended: Permanent AP Red kit, Fast Red substrate kit, New Fuchsin kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out
Procedure:
1. Blocking Solution (protein block, this step is optional) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. AP-polymer anti Mouse 30 min. 6. Washing with wash buffer 3 x 2 min. 7. Fast Red, Permanent AP Red, NBT/BCIP or New Fuchsin 5-15 min. (Controlling the colour intensity via light microscope is recommended.) 8. Stopping the reaction with distilled H2O when the desired colour intensity is attained 9. Counterstaining and blueing 10. Mounting: aqueous with Fast Red, permanent with Permanent AP Red, NBT/BCIP or New Fuchsin
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use a Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus HRP Polymer anti-Rabbit is designed for the qualitative detection of antigens in fixed paraffinembedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbits. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Polymer anti-Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of horse radish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The reagent is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP Polymer anti-Rabbit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRP polymer is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the HRP-polymer is applied and incubated. Any excess of unbound HRP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
100 ml HRP-Polymer anti-Rabbit (Ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC Single Solution, AEC Substrate kit, DAB Substrate kit, DAB High contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. HRP-polymer anti-Rabbit 30 min. 8. Washing with wash buffer 3 x 2 min. 9. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use a Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagent. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus HRP Polymer anti-Rabbit is designed for the qualitative detection of antigens in fixed paraffinembedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbits. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Polymer anti-Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of horse radish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The reagent is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP Polymer anti-Rabbit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRP polymer is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the HRP-polymer is applied and incubated. Any excess of unbound HRP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
6 ml HRP-Polymer anti-Rabbit (Ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC Single Solution, AEC Substrate kit, DAB Substrate kit, DAB High contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. HRP-polymer anti-Rabbit 30 min. 8. Washing with wash buffer 3 x 2 min. 9. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use a Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagent. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus AP Polymer anti-Rabbit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with monoand polyclonal primary antibodies and sera obtained from rabbits. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Polymer anti-Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The reagent is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus AP Polymer anti-Rabbit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP polymer is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the AP-polymer is applied and incubated. Any excess of unbound AP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent AP Red leads to the formation of a magentared product of reaction at the place of the target antigen.
Reagents provided:
6 ml AP-Polymer anti-Rabbit (Ready-to-use) Substrate systems recommended: Permanent AP Red kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. ? Deparaffinise and rehydrate paraffin-embedded tissue sections. ? Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. ? Tissue sections have to be completely covered with the different reagents in order to avoid drying out
Procedure:
1. Blocking Solution (protein block, this step is optional) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. AP-polymer anti Rabbit 30 min. 6. Washing with wash buffer 3 x 2 min. 7. Permanent AP Red 5-15 min. (Controlling the colour intensity via light microscope is recommended.) 8. Stopping the reaction with distilled H2O when the desired colour intensity is attained 9. Counterstaining and blueing 10. Mounting: permanent or aqueous with Permanent AP Red
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use a Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
Aqueous Mount is an aqueous mounting medium for histological and cytological preparations. It is especially suitable for mounting of tissue sections stained with alcohol-soluble chromogenic substrates such as Fast Red or AEC. Even sections stained with alcohol-resistant chromogenic substrates can be mounted with Aqueous Mount if they were not previously dehydrated. Aqueous Mount is intended for research use only.
Aqueous Mount in the following formats: 60 ml Ready-to-use
Storage and handling:
The solution should be stored at room temperature in the original container without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use
Procedure:
Apply a sufficient amount of Aqueous Mount onto the stained histological or cytological section; depending on the size of section 2 to 5 drops. Cover with a coverslip. Heating of the mounted section for hardening is not necessary. Coverslips can be removed by soaking the section over night in water.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. In case of swallowing rinse the mouth excessively with large amounts of water and turn to a doctor. A material safety data sheet (MSDS) is available upon request.
Aqueous Mount is an aqueous mounting medium for histological and cytological preparations. It is especially suitable for mounting of tissue sections stained with alcohol-soluble chromogenic substrates such as Fast Red or AEC. Even sections stained with alcohol-resistant chromogenic substrates can be mounted with Aqueous Mount if they were not previously dehydrated. Aqueous Mount is intended for research use only.
Aqueous Mount in the following formats: 30 ml Ready-to-use
Storage and handling:
The solution should be stored at room temperature in the original container without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use
Procedure:
Apply a sufficient amount of Aqueous Mount onto the stained histological or cytological section; depending on the size of section 2 to 5 drops. Cover with a coverslip. Heating of the mounted section for hardening is not necessary. Coverslips can be removed by soaking the section over night in water.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. In case of swallowing rinse the mouth excessively with large amounts of water and turn to a doctor. A material safety data sheet (MSDS) is available upon request.
Permanent AP Red Kit is developed for immunohistochemical and in situ-hybridisation staining procedures with alkaline phosphatase. Permanent AP Red leads to the formation of a magenta-red precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light or fluorescence microscopy.
500 ml Permanent AP Red Buffer 8 ml Permanent AP Red Chromogen 1 Dilution Vial
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution prepared is stable for about 60 minutes and should therefore be used directly after preparation. Excess working solution should be discarded. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Reagent preparation (Preparation of the working solution) 1) Pipette 2.5 ml AP Red Buffer into the provided dilution vial and let it come to room temperature. The chromogen should still be kept cool. 2) Directly prior to use add 1 drop of Permanent AP Red Chromogen into the buffer. Mix thoroughly. 3) The solution is stable for about 60 minutes. Preparation should be done directly before use. Make sure to pipet the chromogen/substrate mix on the last slide of the staining run within 40 min after mixing. If you want to prepare other quantities of the working solution, please use same ratio AP Red Buffer and Chromogen
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply freshly prepared Permanent AP Red working solution onto the slide. Incubate for 10 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium. Note: It is also possible to mount Permanent AP Red with aqueous mounting media.
Expected results:
During the reaction of the substrate with alkaline phosphatase in presence of the chromogen Permanent AP Red, a magenta-red precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light or fluorescence microscopy (Texas Red filter).
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous alkaline phosphatase activity may cause non-specific staining. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Therefore, tissues of this origin should be stained with peroxidase detection systems. A higher sensitivity can be obtained when a second chromogenic substrate step is used (i. e. 2 x 10 min Permanent AP Red). Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. A longer exposure to absolute ethanol can result in decreasing staining intensity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. A material safety data sheet (MSDS) is available upon request.
Permanent AP Red Kit is developed for immunohistochemical and in situ-hybridisation staining procedures with alkaline phosphatase. Permanent AP Red leads to the formation of a magenta-red precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light or fluorescence microscopy.
125 ml Permanent AP Red Buffer 2 ml Permanent AP Red Chromogen 1 Dilution Vial
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution prepared is stable for about 60 minutes and should therefore be used directly after preparation. Excess working solution should be discarded. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Reagent preparation (Preparation of the working solution) 1) Pipette 2.5 ml AP Red Buffer into the provided dilution vial and let it come to room temperature. The chromogen should still be kept cool. 2) Directly prior to use add 1 drop of Permanent AP Red Chromogen into the buffer. Mix thoroughly. 3) The solution is stable for about 60 minutes. Preparation should be done directly before use. Make sure to pipet the chromogen/substrate mix on the last slide of the staining run within 40 min after mixing. If you want to prepare other quantities of the working solution, please use same ratio AP Red Buffer and Chromogen
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply freshly prepared Permanent AP Red working solution onto the slide. Incubate for 10 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium. Note: It is also possible to mount Permanent AP Red with aqueous mounting media.
Expected results:
During the reaction of the substrate with alkaline phosphatase in presence of the chromogen Permanent AP Red, a magenta-red precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light or fluorescence microscopy (Texas Red filter).
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous alkaline phosphatase activity may cause non-specific staining. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Therefore, tissues of this origin should be stained with peroxidase detection systems. A higher sensitivity can be obtained when a second chromogenic substrate step is used (i. e. 2 x 10 min Permanent AP Red). Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. A longer exposure to absolute ethanol can result in decreasing staining intensity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. A material safety data sheet (MSDS) is available upon request.
DAB Substrate kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). DAB (3,3-Diaminobenzidine) leads to the formation of a brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
30 ml DAB Chromogen (liquid DAB concentrate) 500 ml DAB Substrate Buffer
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Once the two reagents are combined, the resulting solution can be used for up to six hours. Excess working solution needs to be disposed as hazardous substance. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Add 50 µl DAB Chromogen (DAB concentrate) to 1 ml of DAB Substrate Buffer and mix thoroughly. Note: Typical working concentrations are 50 µl (0.9 mg) DAB per ml substrate buffer. The colour intensity can be adjusted by decreasing or increasing the DAB concentration in the working solution. Maximum sensitivity in immunohistochemical staining can be achieved by working concentrations of about 80 µl (1.5 mg) DAB per ml substrate buffer.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the DAB working solution onto the slide. Incubate for 5-15 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium. Note: It is also possible to mount DAB with aqueous mounting media.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen DAB, a brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. The DAB chromogen is hazardous to your health. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
DAB Substrate kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). DAB (3,3-Diaminobenzidine) leads to the formation of a brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
3 ml DAB Chromogen (liquid DAB concentrate) 11 x 5 ml DAB Substrate Buffer
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Once the two reagents are combined, the resulting solution can be used for up to six hours. Excess working solution needs to be disposed as hazardous substance. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Add 4 drops of DAB Chromogen (DAB concentrate) to one bottle of DAB Substrate Buffer and mix thoroughly.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the DAB working solution onto the slide. Incubate for 5-15 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium. Note: It is also possible to mount DAB with aqueous mounting media.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen DAB, a brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. The DAB chromogen is hazardous to your health. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
DAB Substrate kit High Contrast is developed for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). DAB (3,3-Diaminobenzidine) leads to the formation of a brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy. The kit is especially useful when a high contrast between chromogen and counter stain is desired. Compared to standard DAB staining systems the DAB Substrate High Contrast kit gives a darker brown colour and a higher sensitivity.
30 ml DAB Chromogen (liquid DAB concentrate) 500 ml DAB Substrate Buffer High Contrast
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Once the two reagents are combined, the resulting solution is stable for up to six hours. Excess working solution should be disposed as hazardous substance. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Add 50 µl of DAB Chromogen (DAB concentrate) to 1 ml of DAB Substrate Buffer High Contrast and mix thoroughly. Note: Typical working concentrations are 50 µl (0.9 mg) DAB per ml substrate buffer. The colour intensity can be adjusted by decreasing or increasing the DAB concentration in the working solution. Maximum sensitivity in immunohistochemical staining can be achieved by working concentrations of about 80 µl (1.5 mg) DAB per ml substrate buffer.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the DAB High contrast working solution to the slide. Incubate for 5-15 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium. Note: It is also possible to mount DAB High Contrast with aqueous mounting media.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen DAB, a brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. The DAB chromogen is hazardous to your health. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
DAB Substrate kit High Contrast is developed for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). DAB (3,3-Diaminobenzidine) leads to the formation of a brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy. The kit is especially useful when a high contrast between chromogen and counter stain is desired. Compared to standard DAB staining systems the DAB Substrate High Contrast kit gives a darker brown colour and a higher sensitivity.
3 ml DAB Chromogen (liquid DAB concentrate) 11 x 5 ml DAB Substrate Buffer High Contrast
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Once the two reagents are combined, the resulting solution is stable for up to six hours. Excess working solution should be disposed as hazardous substance. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Add 5 drops of DAB Chromogen (DAB concentrate) to one bottle of DAB Substrate Buffer High Contrast and mix thoroughly.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the DAB High contrast working solution to the slide. Incubate for 5-15 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium. Note: It is also possible to mount DAB High Contrast with aqueous mounting media.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen DAB, a brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. The DAB chromogen is hazardous to your health. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
Permanent AEC Kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy. Permanent AEC Kit is intended for research use only.
5.5 ml Reagent 1 3 ml Reagent 2 3 ml Reagent 3 (Chromogen) 4.5 ml Reagent 4 (H2O2) 1 Dilution Vial
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Excess working solution should be disposed. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
1) Pipette 5 ml distilled or deionised water into the provided dilution vial. 2) Add 3 drops buffer concentrate (Reagent 1). Mix thoroughly. 3) Add 2 drops Reagent 2. Mix thoroughly. 4) Add 2 drops AEC chromogen (Reagent 3). Mix thoroughly. 5) Add 2 drops H2O2 substrate (Reagent 4). Mix thoroughly. This working solution is stable for at least 16 hours if stored at 2-8°C in a dark place.
Procedure:
1) Apply the Permanent AEC working solution onto the slide. Incubate for 5-15 minutes. (Incubation time can be extended, if desired.) 2) Rinse with distilled or deionised H2O. 3) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 4) Rinse with distilled or deionised H2O. 5) Blueing in tap water for at least 5 minutes. 6) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. A longer exposure to absolute ethanol can result in decreasing staining intensity. Use of recycled alcohol to dehydrate tissue slides after staining is not recommended. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. A material safety data sheet (MSDS) is available upon request.
AEC Substrate kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous mounting media and can be observed by light microscopy.
15 ml AEC Chromogen (liquid AEC concentrate) 500 ml AEC Substrate Buffer
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Once the two reagents are combined, the resulting solution is stable for up to three hours. Excess working solution needs to be disposed as hazardous substance. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Add 20 µl AEC Chromogen (AEC concentrate) to 1 ml of AEC Substrate Buffer and mix thoroughly. Note: The colour intensity can be adjusted by decreasing or increasing the AEC concentration in the working solution.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the AEC working solution onto the slide. Incubate for 5-20 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Mount with an aqueous mounting medium.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. The coloured precipitate formed by AEC is soluble in organic solvents. The tissue sections therefore have to be counterstained with aqueous solutions (e. g. Gills or Mayers haematoxylin) and mounted with aqueous mounting media. The colour intensity of the reaction product can decrease with time, especially when exposed to light. The staining reaction itself can be influenced in the same way when carried out in strong light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Some of the reagents used in this kit are hazardous to your health. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
AEC Substrate kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous mounting media and can be observed by light microscopy.
3 ml AEC Chromogen (liquid AEC concentrate) 11 x 5 ml AEC Substrate Buffer
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Once the two reagents are combined, the resulting solution is stable for up to three hours. Excess working solution needs to be disposed as hazardous substance. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
Add 2 drops of AEC Chromogen (AEC concentrate) to one bottle of AEC Substrate Buffer and mix thoroughly.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the AEC working solution onto the slide. Incubate for 5-20 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Mount with an aqueous mounting medium.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. The coloured precipitate formed by AEC is soluble in organic solvents. The tissue sections therefore have to be counterstained with aqueous solutions (e. g. Gills or Mayers haematoxylin) and mounted with aqueous mounting media. The colour intensity of the reaction product can decrease with time, especially when exposed to light. The staining reaction itself can be influenced in the same way when carried out in strong light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Some of the reagents used in this kit are hazardous to your health. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining may occur. Material safety data sheets (MSDS) are available upon request.
AEC Single Solution is a ready-to-use solution intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous mounting media and can be observed by light microscopy. AEC Single Solution is especially useful when a high sensitivity is desired.
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. AEC Single Solution is a ready-to-use solution. Preparation of a working solution as in other chromogenic substrates is not necessary. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use. AEC Single Solution can be used directly from the refrigerator and should be stored again at 2-8°C aft er use. When using the small package (8 ml MON-APP169) please directly drop from the bottle. When using the 100 ml package (MON_APP?) please transfer up to 8 ml of the AEC Single Solution into one of the provided dropper bottles. The transferred solution is stable for many weeks if stored at 2-8°C. The vo lume required for several staining runs should be transferred so that the 100 ml stock bottle has to be opened only a few times. If you would like to pipette the solution use a clean vial from which you pipette. Remaining quantities should not be filled back into the bottle but disposed as hazardous material.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the AEC Single Solution to the slide. Incubate for 3-6 minutes. (Incubation time can be extended up to 30 minutes, if desired.) 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Mount with an aqueous mounting medium.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. The coloured precipitate formed by AEC is soluble in organic solvents. The tissue sections therefore have to be counterstained with aqueous solutions (e. g. Gills or Mayers haematoxylin) and mounted with aqueous mounting media. The colour intensity of the reaction product can decrease with time, especially when exposed to light. The staining reaction itself can be influenced in the same way when carried out in strong light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. AEC (3-Amino-9-ethylcarbazol) and the solvents used are considered hazardous materials. Material safety data sheets (MSDS) are available upon request. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Oxidising substances, e. g. metals, dust, bacteria or glass devices can influence the stability of AEC Single Solution. Such contaminations have to be avoided. Non-consumed solution needs to be discarded as dangerous substance.
The wash Buffer is designed as washing solution for immunohistochemical and immunocytological staining procedures on slides. Wash Buffer is primarily used with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures. The Wash Buffer is suitable for manually operated and automated immunohistochemical staining.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. Washing away the applied reagents after each incubation step is critical to receive optimally stained samples. The Wash Buffer is especially designed for effective washing and therefore ensures brilliant staining results.
Principle of method:
The Wash Buffer is a 20fold concentrated phosphate buffer with additives of sodium chloride, detergent, and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:20 with deionised or distilled water. The resulting solution has a pH of 7.2 (7.0 to 7.4). The Wash Buffer is wetting the tissue sections with detergent and thus reduces surface tension and improves spreading the reagents on the tissue section, reduces unspecific binding of reagents on the tissue sample, and because of the exact tuned salt concentration effects an excellent preservation of cell morphology.
Reagents provided:
2500 ml Wash Buffer (20fold concentrated, adequate for 50 litres ready-to-use wash buffer)
Storage and handling:
The solution should be stored at room temperature. It is stable up to the expiry date indicated on the label if undiluted. Do not use product after the expiry date. The diluted working strength solution is stable for about 1 week depending on the ambient temperature. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the Wash Buffer working strength solution: Dilute Wash Buffer concentrate 1:20 with deionised or distilled water and mix thoroughly. The pH-value should be at 7.2 (7.0 to 7.4). If necessary adjust pH-value with diluted NaOH or HCl solution.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The Wash Buffer is a 20fold concentrated solution with a mildly acidic pH-value. The correct pHvalue of about 7.0 (+/- 0.2) is achieved after diluting the solution 1:20. Sometimes deionised water has pH-values considerably different from the neutral point (pH 7.0) depending on the preparation method. Experiments have shown that the Wash Buffer can successfully be diluted with deionised or distilled with water in the pH-range of 5.5 up to 9.5. If a detection system with alkaline phosphatase is used please note: larger amounts of wash buffer remaining on the slides can lead to decreasing enzyme activity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
The wash Buffer is designed as washing solution for immunohistochemical and immunocytological staining procedures on slides. Wash Buffer is primarily used with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures. The Wash Buffer is suitable for manually operated and automated immunohistochemical staining.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. Washing away the applied reagents after each incubation step is critical to receive optimally stained samples. The Wash Buffer is especially designed for effective washing and therefore ensures brilliant staining results.
Principle of method:
The Wash Buffer is a 20fold concentrated phosphate buffer with additives of sodium chloride, detergent, and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:20 with deionised or distilled water. The resulting solution has a pH of 7.2 (7.0 to 7.4). The Wash Buffer is wetting the tissue sections with detergent and thus reduces surface tension and improves spreading the reagents on the tissue section, reduces unspecific binding of reagents on the tissue sample, and because of the exact tuned salt concentration effects an excellent preservation of cell morphology.
Reagents provided:
500 ml Wash Buffer (20fold concentrated, adequate for 10 litres ready-to-use wash buffer)
Storage and handling:
The solution should be stored at room temperature. It is stable up to the expiry date indicated on the label if undiluted. Do not use product after the expiry date. The diluted working strength solution is stable for about 1 week depending on the ambient temperature. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the Wash Buffer working strength solution: Dilute Wash Buffer concentrate 1:20 with deionised or distilled water and mix thoroughly. The pH-value should be at 7.2 (7.0 to 7.4). If necessary adjust pH-value with diluted NaOH or HCl solution.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The Wash Buffer is a 20fold concentrated solution with a mildly acidic pH-value. The correct pHvalue of about 7.0 (+/- 0.2) is achieved after diluting the solution 1:20. Sometimes deionised water has pH-values considerably different from the neutral point (pH 7.0) depending on the preparation method. Experiments have shown that the Wash Buffer can successfully be diluted with deionised or distilled with water in the pH-range of 5.5 up to 9.5. If a detection system with alkaline phosphatase is used please note: larger amounts of wash buffer remaining on the slides can lead to decreasing enzyme activity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER T-EDTA Buffer pH 9.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffinembedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consists of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER T-EDTA Buffer pH 9.0 is a 10fold concentrated EDTA solution in Tris buffer with additives of detergent and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 9.0 (8.8 to 9.2). HIER T-EDTA Buffer pH 9.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of many different specificities. It leads to considerably stronger signals compared with usually used citrate buffer.
Reagents provided:
500 ml HIER T-EDTA Buffer pH 9.0 (10fold concentrated, adequate for 5 litres ready-to-use T-EDTA Buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the T-EDTA buffer working strength solution: Dilute HIER T-EDTA Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 9.0 (8.8 to 9.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER T-EDTA Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with T-EDTA buffer in the steamer, close the lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with T-EDTA buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove T-EDTA buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER T-EDTA Buffer pH 9.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffinembedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consists of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER T-EDTA Buffer pH 9.0 is a 10fold concentrated EDTA solution in Tris buffer with additives of detergent and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 9.0 (8.8 to 9.2). HIER T-EDTA Buffer pH 9.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of many different specificities. It leads to considerably stronger signals compared with usually used citrate buffer.
Reagents provided:
100 ml HIER T-EDTA Buffer pH 9.0 (10fold concentrated, adequate for 1 litre ready-to-use T-EDTA Buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the T-EDTA buffer working strength solution: Dilute HIER T-EDTA Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 9.0 (8.8 to 9.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER T-EDTA Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with T-EDTA buffer in the steamer, close the lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with T-EDTA buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove T-EDTA buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER EDTA Buffer pH 8.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffinembedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to the antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER EDTA Buffer pH 8.0 is a 10fold concentrated, buffered EDTA solution with additives of detergent and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 8.0 (7.8 to 8.2). HIER EDTA Buffer pH 8.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of different specificity. HIER EDTA Buffer pH 8.0 leads to considerably stronger signals compared with usually used citrate buffer.
Reagents provided:
500 ml HIER EDTA Buffer pH 8.0 (10fold concentrated, adequate for 5 litres ready-to-use EDTA Buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the EDTA buffer working strength solution: Dilute HIER EDTA Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 8.0 (7.8 to 8.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER EDTA Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with EDTA buffer in the steamer, close lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with EDTA buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove EDTA buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER Citrate Buffer pH 6.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffin-embedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to the antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER Citrate Buffer pH 6.0 is a 10fold concentrated citrate buffer solution with additives of stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 6.0 (5.8 to 6.2). HIER Citrate Buffer pH 6.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of many different specificities.
Reagents provided:
500 ml HIER Citrate Buffer pH 6.0 (10fold concentrated, adequate for 5 litres ready-to-use citrate buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negat ive control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the citrate buffer working strength solution: Dilute HIER Citrate Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 6.0 (5.8 to 6.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER Citrate Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with citrate buffer in the steamer, close the lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with citrate buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove citrate buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER Citrate Buffer pH 6.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffin-embedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to the antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER Citrate Buffer pH 6.0 is a 10fold concentrated citrate buffer solution with additives of stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 6.0 (5.8 to 6.2). HIER Citrate Buffer pH 6.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of many different specificities.
Reagents provided:
100 ml HIER Citrate Buffer pH 6.0 (10fold concentrated, adequate for 1 litre ready-to-use citrate buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negat ive control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the citrate buffer working strength solution: Dilute HIER Citrate Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 6.0 (5.8 to 6.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER Citrate Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with citrate buffer in the steamer, close the lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with citrate buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove citrate buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
Fast Enzyme is a ready-to-use solution developed for enzymatic epitope retrieval on formalin-fixed tissue sections on slides. This procedure (sometimes called PIER, Protease Induced Epitope Retrieval) is primarily used in immunohistochemical staining procedures. Proteolytic pre-treatment is also used in in situ-hybridisation. Fast Enzyme is intended for research use only.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Enzymatic digestion with proteolytic enzymes (PIER) restores structures of the epitopes making them more accessible to specific antibodies. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
Fast Enzyme is a ready-to-use enzyme solution for enzymatic epitope retrieval.
Reagents provided:
15 ml Fast Enzyme (Ready-To-Use)
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
The solution is ready-to-use.
Procedure:
Fast Enzyme is suitable for enzymatic epitope retrieval carried out after the dewaxing and rehydration of the tissue sections. 1. Cover deparaffinised and rehydrated tissue sections with ready-to-use Fast Enzyme Solution. 2. Incubate for 5 minutes at room temperature. (It was shown that in individual cases a stronger signal can be obtained when the incubation time is elongated. Usually an incubation for 5 min at room temperatur is sufficient.) 3. Rinse carefully (3 x) with wash buffer. 4. Proceed with immunohistological staining as usual.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. A material safety data sheet (MSDS) is available upon request.
Trypsin Pretreatment Kit consists of 2 reagents for the preparation of a trypsin solution used for enzymatic epitope retrieval on formalin-fixed tissue sections on slides. This procedure (sometimes called PIER, Protease Induced Epitope Retrieval) is primarily used in immunohistochemical staining procedures. Trypsin Pretreatment Kit is for research use only.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Enzymatic digestion with proteolytic enzymes (PIER) restores structures of the epitopes making them more accessible to specific antibodies. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
The components of this Trypsin Pretreatment Kit allow for the preparation of a buffered trypsin solution for enzymatic epitope retrieval.
Reagents provided:
30 ml Trypsin Solution 125 ml Trypsin Buffer
Storage and handling:
The solutions should be stored at 2-8°C. Please sto re the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Mix 1 part Trypsin Solution with 3 parts Trypsin Buffer. The activity of the resulting trypsin solution can be adjusted by variation of the mixing ratio. Mix the two components in the ratio 1:1 when strong epitope retrieval is desired. The working solution is stable for at least one week if stored at 2-8°C.
Procedure:
Trypsin Pretreatment Kit is suitable for enzymatic epitope retrieval carried out after the dewaxing and rehydration of the sections. 1. Cover deparaffinised and rehydrated tissue sections with trypsin working solution. 2. Incubate for 10 - 20 minutes at 37°C. 3. Rinse carefully (3 x) with wash buffer. 4. Proceed with immunohistological staining as usual.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. A material safety data sheet (MSDS) is available upon request.
Pronase Solution is a ready-to-use solution developed for enzymatic epitope retrieval on formalin-fixed tissue sections on slides. This procedure (sometimes called PIER, Protease Induced Epitope Retrieval) is primarily used in immunohistochemical staining procedures. Proteolytic pre-treatments are also used in protocols for in situhybridization. Pronase solution is intended for research use only.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Enzymatic digestion with proteolytic enzymes (PIER) restores structures of the epitopes making them more accessible to specific antibodies. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
Pronase Solution is a ready-to-use solution for enzymatic epitope retrieval.
Reagents provided:
250 ml Pronase Solution (Ready-To-Use)
Storage and handling:
The solution should be stored in aliquots at -20°C without further dilution. The solution should be aliquoted in order to avoid repeated freeze and thawing. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date.
Reagent preparation:
Pronase solution is ready-to-use and should be at room temperature prior to use.
Procedure:
Pronase Solution is suitable for enzymatic epitope retrieval carried out after the dewaxing and rehydration of the tissue sections. 1. Cover deparaffinised and rehydrated tissue sections with ready-to-use Pronase Solution. 2. Incubate for 15 - 20 minutes at room temperature. The optimal incubation time needs to be elaborated by the operator. 3. Rinse carefully (3 x), first with distilled water followed by buffer. 4. Proceed with immunohistological staining as usual.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of the reagent with eye, skin or mucous membrane. In case of reagent coming into contact with a sensitive area, wash the area with large amounts of water. Material safety data sheets (MSDS) are available upon request.
Pepsin Solution is a ready-to-use solution developed for enzymatic epitope retrieval on formalin-fixed tissue sections on slides. This procedure (sometimes called PIER, Protease Induced Epitope Retrieval) is primarily used in immunohistochemical staining procedures.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Enzymatic digestion with proteolytic enzymes (PIER) restores structures of the epitopes making them more accessible to specific antibodies. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
Pepsin Solution is a ready-to-use stabilised pepsin solution for enzymatic epitope retrieval.
Reagents provided:
60 ml Pepsin Solution (Ready-To-Use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support
Reagent preparation:
The solution is ready-to-use.
Procedure:
Pepsin Solution is suitable for enzymatic epitope retrieval carried out after the dewaxing and rehydration of the tissue sections. 1. Cover deparaffinised and rehydrated tissue sections with ready-to-use Pepsin Solution. 2. Incubate for 10 - 15 minutes at 37°C or 20 30 minutes at room temperature. The optimal incubation time needs to be elaborated by the operator. It was shown that in individual cases a stronger signal can be obtained when the incubation time is elongated up to 120 minutes (e. g. for detection of Collagen IV with different antibodies). 3. Rinse carefully (3 x) with wash buffer. 4. Proceed with immunohistological staining as usual.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. A material safety data sheet (MSDS) is available upon request.
Antibody Diluent B is especially developed for dilution of certain primary antibodies. Antibodies diluted with Antibody Diluent B are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent B reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Principle of method:
Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent B. Antibody Diluent B minimises unspecific binding of the primary antibody to the tissue section, reduces surface tension of the antibody solution and improves spreading the reagent on the slide, increases microbial and chemical stability of the antibody, reduces adhesion of antibody to the surface of the vial, and minimises the danger of antibody degradation by proteolytic enzymes.
Reagents provided:
100 ml Antibody Diluent B (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results.Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. (NaN3), used for stabilisation, is not considered hazardous material in the concentration used. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. A material safety data sheet (MSDS) is available upon request.
Antibody Diluent B is especially developed for dilution of certain primary antibodies. Antibodies diluted with Antibody Diluent B are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent B reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Principle of method:
Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent B. Antibody Diluent B minimises unspecific binding of the primary antibody to the tissue section, reduces surface tension of the antibody solution and improves spreading the reagent on the slide, increases microbial and chemical stability of the antibody, reduces adhesion of antibody to the surface of the vial, and minimises the danger of antibody degradation by proteolytic enzymes.
Reagents provided:
25 ml Antibody Diluent B (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results.Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. (NaN3), used for stabilisation, is not considered hazardous material in the concentration used. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. A material safety data sheet (MSDS) is available upon request.
Antibody Diluent is developed for dilution of primary antibodies. Antibodies diluted with Antibody Diluent are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Principle of method:
Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent. Antibody Diluent ? minimises unspecific binding of the primary antibody to the tissue section, ? reduces surface tension of the antibody solution and improves spreading the reagent on the slide, ? increases microbial and chemical stability of the antibody, ? reduces adhesion of antibody to the surface of the vial, ? and minimises the danger of antibody degradation by proteolytic enzymes.
Reagents provided:
500 ml Antibody Diluent (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use through qualified personnel only. Wear protective clothing to avoid contact of reagents and specimens with eye, skin and mucous membranes. If reagents or specimens come in contact with sensitive area, wash with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin300 and sodium azide (NaN3) are used for stabilisation. Reaction of sodium azide with lead or copper in drainage pipes can result in the formation of highly explosive metallic azides. Discard the antibody solution in a large volume of running water to avoid formation of deposits. A material safety data sheet (MSDS) for the pure substances is available upon request.
Antibody Diluent is developed for dilution of primary antibodies. Antibodies diluted with Antibody Diluent are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Principle of method:
Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent. Antibody Diluent ? minimises unspecific binding of the primary antibody to the tissue section, ? reduces surface tension of the antibody solution and improves spreading the reagent on the slide, ? increases microbial and chemical stability of the antibody, ? reduces adhesion of antibody to the surface of the vial, ? and minimises the danger of antibody degradation by proteolytic enzymes.
Reagents provided:
100 ml Antibody Diluent (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use through qualified personnel only. Wear protective clothing to avoid contact of reagents and specimens with eye, skin and mucous membranes. If reagents or specimens come in contact with sensitive area, wash with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin300 and sodium azide (NaN3) are used for stabilisation. Reaction of sodium azide with lead or copper in drainage pipes can result in the formation of highly explosive metallic azides. Discard the antibody solution in a large volume of running water to avoid formation of deposits. A material safety data sheet (MSDS) for the pure substances is available upon request.
The blocking solution Peroxide Block is intended for inhibition of endogenous peroxidase activity in tissue sections. It is primarily intended to be used in immunohistochemistry on formalin-fixed paraffin-embedded samples if using a detection system with horse radish peroxidase.
Endogenous peroxidase activity in the tissue can result in unspecific background staining in immunohistochemical staining procedures using horse radish peroxidase (HRP) as detection enzyme. This effect can be eliminated when tissue sections are incubated with Peroxide Block prior to immunohistochemical staining. Hydrogen peroxide in the solution blocks the activity of endogenous peroxidase.
Principle of method:
Peroxide Block is applied onto tissue sections to reduce non-specific staining due to endogenous peroxidase activity in immunohistochemistry. The step is carried out before incubation with primary antibody but after dewaxing and rehydration. If a heat induced epitope retrieval (HIER) or enzymatic digestion is necessary for immunohistochemical detection it is of no importance if the Peroxide Block is used before or after this step. In some cases it has been shown, that blocking of endogenous peroxidase before the epitope retrieval leads to better results.
Reagents provided:
500 ml Peroxide Block (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Avoid exposure to strong light. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Procedure:
1. Apply Peroxide Block for 10 minutes at room temperature. The section should be covered completely. 2. Rinse with wash buffer. 3. Proceed with next steps for immunohistochemical staining as usual starting with protein blocking or the primary antibody.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. A material safety data sheet (MSDS) is available upon request
The blocking solution Peroxide Block is intended for inhibition of endogenous peroxidase activity in tissue sections. It is primarily intended to be used in immunohistochemistry on formalin-fixed paraffin-embedded samples if using a detection system with horse radish peroxidase.
Endogenous peroxidase activity in the tissue can result in unspecific background staining in immunohistochemical staining procedures using horse radish peroxidase (HRP) as detection enzyme. This effect can be eliminated when tissue sections are incubated with Peroxide Block prior to immunohistochemical staining. Hydrogen peroxide in the solution blocks the activity of endogenous peroxidase.
Principle of method:
Peroxide Block is applied onto tissue sections to reduce non-specific staining due to endogenous peroxidase activity in immunohistochemistry. The step is carried out before incubation with primary antibody but after dewaxing and rehydration. If a heat induced epitope retrieval (HIER) or enzymatic digestion is necessary for immunohistochemical detection it is of no importance if the Peroxide Block is used before or after this step. In some cases it has been shown, that blocking of endogenous peroxidase before the epitope retrieval leads to better results.
Reagents provided:
100 ml Peroxide Block (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Avoid exposure to strong light. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Procedure:
1. Apply Peroxide Block for 10 minutes at room temperature. The section should be covered completely. 2. Rinse with wash buffer. 3. Proceed with next steps for immunohistochemical staining as usual starting with protein blocking or the primary antibody.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. A material safety data sheet (MSDS) is available upon request
The blocking solution Peroxide Block is intended for inhibition of endogenous peroxidase activity in tissue sections. It is primarily intended to be used in immunohistochemistry on formalin-fixed paraffin-embedded samples if using a detection system with horse radish peroxidase.
Endogenous peroxidase activity in the tissue can result in unspecific background staining in immunohistochemical staining procedures using horse radish peroxidase (HRP) as detection enzyme. This effect can be eliminated when tissue sections are incubated with Peroxide Block prior to immunohistochemical staining. Hydrogen peroxide in the solution blocks the activity of endogenous peroxidase.
Principle of method:
Peroxide Block is applied onto tissue sections to reduce non-specific staining due to endogenous peroxidase activity in immunohistochemistry. The step is carried out before incubation with primary antibody but after dewaxing and rehydration. If a heat induced epitope retrieval (HIER) or enzymatic digestion is necessary for immunohistochemical detection it is of no importance if the Peroxide Block is used before or after this step. In some cases it has been shown, that blocking of endogenous peroxidase before the epitope retrieval leads to better results.
Reagents provided:
8 ml Peroxide Block (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Avoid exposure to strong light. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Procedure:
1. Apply Peroxide Block for 10 minutes at room temperature. The section should be covered completely. 2. Rinse with wash buffer. 3. Proceed with next steps for immunohistochemical staining as usual starting with protein blocking or the primary antibody.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. A material safety data sheet (MSDS) is available upon request
Blocking Solution is developed to eliminate unspecific binding of primary and secondary antibodies to tissue sections. It is primarily intended to be used in immunohistochemistry on formalin-fixed paraffin-embedded samples.
Unspecific binding of primary and secondary antibodies to tissue sections in immunohistochemical staining procedures can result in background staining. This effect can be eliminated when tissue sections are incubated with Blocking Solution prior to incubation with the primary antibody. The protein in Blocking Solution abolishes unspecific binding. Blocking Solution is a universal blocking reagent. Unlike other frequently used blocking solutions (e. g. serum blocks) the reagent can be used regardless of the origin species of the secondary antibody. Interferences with secondary antibodies or other components of detection systems are not observed. In contrast to other blocking reagents this Blocking Solution should not be incubated longer than 10 minutes and should be rinsed away with wash buffer. Otherwise the signal intensity of the immunohistochemical staining reaction could be decreased.
Principle of method:
Blocking Solution is applied on tissue sections to reduce background staining due to unspecific binding of primary and secondary antibodies in immunohistochemistry. The step is carried out prior to incubation with the primary antibody
Reagents provided:
100 ml Blocking Solution (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Procedure:
1. Apply Blocking Solution for 5 minutes at room temperature. The section should be covered completely. 2. Rinse with wash buffer. 3. Proceed with next steps for immunohistochemical staining as usual starting with the primary antibody.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support . Also refer to the instructions of the detection systems containing Blocking Solution for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 950 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus HRP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus HRP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRPconjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP132) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP133) forms a dark brown precipitate.
Reagents provided:
4x 15 ml Blocking Solution Reagent 1 (ready-to-use) 4x 15 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 4x 15 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC Single Solution, AEC substrate kit, DAB Substrate kit, DAB High Contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Procedure:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP132 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP133 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus HRP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRPconjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP132) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP133) forms a dark brown precipitate.
Reagents provided:
500 ml Blocking Solution Reagent 1 (ready-to-use) 500 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 500 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC Single Solution, AEC substrate kit, DAB Substrate kit, DAB High Contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Procedure:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP132 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP133 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus HRP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRPconjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP132) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP133) forms a dark brown precipitate.
Reagents provided:
125 ml Blocking Solution Reagent 1 (ready-to-use) 125 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 125 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC Single Solution, AEC substrate kit, DAB Substrate kit, DAB High Contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Procedure:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP132 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP133 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus HRP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRPconjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP132) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP133) forms a dark brown precipitate.
Reagents provided:
8 ml Peroxide Block (ready-to-use) 8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 8 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) 7 x 5 ml DAB Substrate Buffer 3 ml DAB Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC Single Solution, AEC substrate kit, DAB Substrate kit, DAB High Contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Procedure:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP132 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP133 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus HRP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRPconjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP132) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP133) forms a dark brown precipitate.
Reagents provided:
8 ml Peroxide Block (ready-to-use) 8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 8 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) 7 x 5 ml AEC Substrate Buffer 3 ml AEC Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC Single Solution, AEC substrate kit, DAB Substrate kit, DAB High Contrast kit Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Procedure:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP132 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP133 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus AP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus AP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP128) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
4x 15 ml Blocking Solution Reagent 1 (ready-to-use) 4x 15 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 4x 15 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit) Permanent AP Red kit, BCIP/NBT Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP128 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Rabbit (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus AP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus AP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP128) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
500 ml Blocking Solution Reagent 1 (ready-to-use) 500 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 500 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit) Permanent AP Red kit, BCIP/NBT Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP128 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Rabbit (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus AP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus AP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP128) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
125 ml Blocking Solution Reagent 1 (ready-to-use) 125 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 125 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit) Permanent AP Red kit, BCIP/NBT Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP128 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Rabbit (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus AP Kit, Rabbit is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from rabbit. The Plus AP Kit, Rabbit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Rabbit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Rabbit binds to rabbit primary antibodies. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from rabbit.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP128) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, Rabbit Reagent 2 (ready-to-use) 8 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) 8 x 5 ml Permanent Red Buffer (Substrate Buffer) 2 ml Permanent Red Concentrate (Chromogen) Substrate systems recommended (if not included in the kit) Permanent AP Red kit, BCIP/NBT Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP128 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Rabbit (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with AP008RED-RB) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus HRP Kits, Mouse is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus HRP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed via a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP123) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP124) forms a dark brown precipitate.
Reagents provided:
4x 15 ml Blocking Solution Reagent 1 (ready-to-use) 4x 15 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 4x 15 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP123 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP124 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kits, Mouse is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus HRP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed via a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP123) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP124) forms a dark brown precipitate.
Reagents provided:
500 ml Blocking Solution Reagent 1 (ready-to-use) 500 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 500 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP123 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP124 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kits, Mouse is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus HRP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed via a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP123) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP124) forms a dark brown precipitate.
Reagents provided:
125 ml Blocking Solution Reagent 1 (ready-to-use) 125 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 125 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP123 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP124 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kits, Mouse is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus HRP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed via a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP123) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP124) forms a dark brown precipitate.
Reagents provided:
8 ml Peroxide Block (ready-to-use) 8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 8 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) 7 x 5 ml DAB Substrate Buffer 3 ml DAB Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP123 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP124 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kits, Mouse is based on the streptavidin-biotin system. It is designed for qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus HRP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed via a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP123) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP124) forms a dark brown precipitate.
Reagents provided:
8 ml Peroxide Block (ready-to-use) 8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 8 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) 7 x 5 ml AEC Substrate Buffer 3 ml AEC Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP123 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP124 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus AP Kit, Mouse is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus AP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidinalkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP119) leads to a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
4x 15 ml Blocking Solution Reagent 1 (ready-to-use) 4x 15 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 4x 15 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP119 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear
The Plus AP Kit, Mouse is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus AP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidinalkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP119) leads to a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
500 ml Blocking Solution Reagent 1 (ready-to-use) 500 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 500 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP119 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear
The Plus AP Kit, Mouse is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus AP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidinalkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP119) leads to a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
125 ml Blocking Solution Reagent 1 (ready-to-use) 125 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 125 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP119 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear
The Plus AP Kit, Mouse is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The Plus AP Kit, Mouse can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kit, Mouse is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Mouse binds to mouse primary antibodies. Therefore this kit can detect monoclonal primary antibodies and sera obtained from mice.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidinalkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP119) leads to a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, Mouse Reagent 2 (ready-to-use) 8 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) 8 x 5 ml Permanent Red Buffer (Substrate Buffer) 2 ml Permanent Red Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP119 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (Substrate Buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP119) 5 min. 12. Wash with destilled water 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear
The Plus HRP Kit, Broad Spectrum is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig. The Plus HRP Kit, Broad Spectrum can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Broad Spectrum is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Broad Spectrum is polyvalent. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP114) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP115) forms a dark brown precipitate.
Reagents provided:
8 ml Peroxide Block (ready-to-use) 8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, polyvalent Reagent 2 (ready-to-use) 8 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) 7 x 5 ml DAB Substrate Buffer 3 ml DAB Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP114 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP115 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, polyvalent (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, rabbit, rat or guinea pig. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus HRP Kit, Broad Spectrum is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig. The Plus HRP Kit, Broad Spectrum can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Kit, Broad Spectrum is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and horse radish peroxidase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus HRP Kit, Broad Spectrum is polyvalent. Therefore this kit can detect mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (Peroxide Block). Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (Blocking Solution). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidin-horse radish peroxidase-conjugate (Streptavidin-HRP-Conjugate). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-HRP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the horse radish peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC (included only in kit MON-APP114) leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB (included only in kit MON-APP115) forms a dark brown precipitate.
Reagents provided:
8 ml Peroxide Block (ready-to-use) 8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, polyvalent Reagent 2 (ready-to-use) 8 ml Streptavidin-HRP-Conjugate Reagent 3 (ready-to-use) 7 x 5 ml AEC Substrate Buffer 3 ml AEC Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB high contrast kit. Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. The tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate AEC working solution (with MON-APP114 only): Add 2 drops (100 µl) of AEC Concentrate to one bottle of AEC Substrate Buffer and mix thoroughly. Preparation of the chromogenic substrate DAB working solution (with MON-APP115 only): Add 4 drops (200 µl) of DAB Concentrate to one bottle of DAB Substrate Buffer and mix thoroughly.
Procedure:
1. Peroxide Block (3% H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 2 min. 7. Biotinylated Secondary Antibody, polyvalent (Reagent 2, yellow) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Streptavidin-HRP-Conjugate (Reagent 3, red) 10-15 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, rabbit, rat or guinea pig. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase. Maybe the hydrogen peroxide solution used for blocking was inactivated. 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with 3% H2O2 solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Background staining due to endogenous biotin can be blocked through an avidinbiotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and sodium azide (NaN3) are used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) are available upon request.
The Plus AP Kit, Broad Spectrum is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig. The Plus AP Kit, Broad Spectrum can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kits, Broad Spectrum is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Broad Spectrum is polyvalent. With this kit it is therefore possible to detect mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized via incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP110) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
4x 15 ml Blocking Solution Reagent 1 (ready-to-use) 4x 15 ml Biotinylated Secondary Antibody, polyvalent Reagent 2 (ready-to-use) 4x 15 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) 8 x 5 ml Permanent Red Buffer (Substrate Buffer) 2 ml Permanent Red Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion.Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP110 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (substrate buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, polyvalent (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 12. Wash with distilled H2O 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, rabbit, rat or guinea pig. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolized by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see also Limitations of the procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3), used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus AP Kit, Broad Spectrum is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig. The Plus AP Kit, Broad Spectrum can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kits, Broad Spectrum is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Broad Spectrum is polyvalent. With this kit it is therefore possible to detect mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized via incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP110) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
500 ml Blocking Solution Reagent 1 (ready-to-use) 500 ml Biotinylated Secondary Antibody, polyvalent Reagent 2 (ready-to-use) 500 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion.Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP110 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (substrate buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, polyvalent (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 12. Wash with distilled H2O 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, rabbit, rat or guinea pig. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolized by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see also Limitations of the procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3), used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus AP Kit, Broad Spectrum is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig. The Plus AP Kit, Broad Spectrum can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kits, Broad Spectrum is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Broad Spectrum is polyvalent. With this kit it is therefore possible to detect mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized via incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP110) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
125 ml Blocking Solution Reagent 1 (ready-to-use) 125 ml Biotinylated Secondary Antibody, polyvalent Reagent 2 (ready-to-use) 125 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion.Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP110 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (substrate buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, polyvalent (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 12. Wash with distilled H2O 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, rabbit, rat or guinea pig. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolized by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see also Limitations of the procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3), used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus AP Kit, Broad Spectrum is based on the streptavidin-biotin system. It is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. The kit is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig. The Plus AP Kit, Broad Spectrum can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Kits, Broad Spectrum is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The method is based on the streptavidin-biotin system which means that a biotinylated secondary antibody binds to several molecules of a conjugate composed of streptavidin and alkaline phosphatase. Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The biotinylated secondary antibody in the Plus AP Kit, Broad Spectrum is polyvalent. With this kit it is therefore possible to detect mono- and polyclonal primary antibodies and sera obtained from mouse, rabbit, rat, and guinea pig.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized via incubation with a protein blocking solution (Blocking Solution provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and streptavidin-alkaline phosphatase-conjugate (Streptavidin-AP-Conjugate). A second washing is followed by the application of this conjugate. It binds to biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent Red (included only in kit MON-APP110) leads to the formation of a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red) or NBT (blue-black) with its substrate BCIP.
Reagents provided:
8 ml Blocking Solution Reagent 1 (ready-to-use) 8 ml Biotinylated Secondary Antibody, polyvalent Reagent 2 (ready-to-use) 8 ml Streptavidin-AP-Conjugate Reagent 3 (ready-to-use) 8 x 5 ml Permanent Red Buffer (Substrate Buffer) 2 ml Permanent Red Concentrate (Chromogen) Substrate systems recommended (if not included in the kit): Permanent AP Red Kit, BCIP/NBT Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion.Tissue sections have to be completely covered with the different reagents in order to avoid drying out. Preparation of the chromogenic substrate working solution (with MON-APP110 only): Add 2 drops (60 µl) of Permanent Red Concentrate to one bottle of Permanent Red Buffer (substrate buffer) and mix. This solution should be used directly after preparation.
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, polyvalent (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 10. Wash with distilled H2O 1 min. 11. Permanent Red substrate-chromogen solution (with MON-APP110) 5 min. 12. Wash with distilled H2O 3 x 1 min. 13. Counterstaining and blueing 14. Mounting: aqueous or permanent after dehydration * The incubation times should be adjusted, when using other substrate-chromogen systems.
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse, rabbit, rat or guinea pig. 6. The antigen was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme conjugate or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolized by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see also Limitations of the procedure). 9. Non-specific binding of the secondary antibody to endogenous biotin in the tissue section. Carry out an avidin-biotin block before incubation with the primary antibody.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity or the endogenous biotin content may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. ProClin 300 and sodium azide (NaN3), used for stabilisation. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. Material safety data sheets (MSDS) for the pure substances are available upon request. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear.
The Plus HRP One-Step Polymer anti-Mouse/Rabbit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It was developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP One-Step Polymer anti-Mouse/Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer consists of several molecules of secondary antibodies covalently bound to several molecules of horseradish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP One-Step Polymer kits avoid the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRP One-Step Polymer is minimized by incubation with a protein blocking solution. This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the HRP One-Step Polymer is applied and incubated. Any excess of unbound polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
100 mL HRP One-Step-Polymer anti-Mouse/Rabbit (ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB High contrast kit. Materials required but not supplied: Positive and negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. HRP One-Step Polymer anti-Mouse/Rabbit 30 min. 8. Washing with wash buffer 3 x 2 min. 9. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with AEC, permanent with DAB
Expected results:
During the reaction of the substrate with horseradish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horseradish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution can result in decreasing signal intensity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus HRP One-Step Polymer anti-Mouse/Rabbit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It was developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP One-Step Polymer anti-Mouse/Rabbit is a highly sensitive detection reagent intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer consists of several molecules of secondary antibodies covalently bound to several molecules of horseradish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice or rabbit. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP One-Step Polymer kits avoid the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRP One-Step Polymer is minimized by incubation with a protein blocking solution. This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the HRP One-Step Polymer is applied and incubated. Any excess of unbound polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
6 mL HRP One-Step-Polymer anti-Mouse/Rabbit (ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB High contrast kit. Materials required but not supplied: Positive and negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. HRP One-Step Polymer anti-Mouse/Rabbit 30 min. 8. Washing with wash buffer 3 x 2 min. 9. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with AEC, permanent with DAB
Expected results:
During the reaction of the substrate with horseradish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use Blocking Solution or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horseradish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution can result in decreasing signal intensity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheet (MSDS) is available upon request.
The Plus HRP Polymer anti-Mouse kit is designed for the qualitative detection of antigens in fixed paraffinembedded tissue sections, in frozen tissue sections, and in cytological samples. It was developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Polymer anti-Mouse kit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer consists of several molecules of secondary antibodies covalently bound to several molecules of horse radish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of monoclonal primary antibodies and sera obtained from mice. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP Polymer antiMouse kit avoids the problem of background staining caused by endogenous biotin in the tissue
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRPpolymer is minimized by incubation with a protein blocking solution. This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the HRP-polymer is applied and incubated. Any excess of unbound HRP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
100 mL HRP-Polymer anti-Mouse (ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB High contrast kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagent should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. HRP-polymer anti-Mouse 30 min. 8. Washing with wash buffer 3 x 2 min. 9. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution can result in decreasing signal intensity. Sanbio guarantee that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheets (MSDS) is available upon request.
The Plus HRP Polymer anti-Mouse kit is designed for the qualitative detection of antigens in fixed paraffinembedded tissue sections, in frozen tissue sections, and in cytological samples. It was developed for use in combination with monoclonal primary antibodies and sera obtained from mice. The reagent can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Polymer anti-Mouse kit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer consists of several molecules of secondary antibodies covalently bound to several molecules of horse radish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of monoclonal primary antibodies and sera obtained from mice. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP Polymer antiMouse kit avoids the problem of background staining caused by endogenous biotin in the tissue
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRPpolymer is minimized by incubation with a protein blocking solution. This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the HRP-polymer is applied and incubated. Any excess of unbound HRP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
6 mL HRP-Polymer anti-Mouse (ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB High contrast kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date. It should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support .
Reagent preparation:
Reagent should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. HRP-polymer anti-Mouse 30 min. 8. Washing with wash buffer 3 x 2 min. 9. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with AEC, permanent with DAB or Permanent AEC
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution can result in decreasing signal intensity. Sanbio guarantee that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of the reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining might appear. ProClin 950 is used for stabilisation. A Material safety data sheets (MSDS) is available upon request.
The Plus HRP Polymer Kit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Polymer Kit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of horse radish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP Polymer Kit avoids the problem of background staining caused by endogenous biotin in the tissue
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRPpolymer is minimized by incubation with a protein blocking solution (Blocking Solution, provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the enhancement reagent (PostBlock) is applied and incubated. A second washing is followed by the application of the HRP-polymer. Any excess of unbound HRP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
100 ml Blocking Solution Reagent 1 (ready-to-use) 100 ml PostBlock Reagent 2 (ready-to-use) 100 ml HRP-Polymer (Mouse/Rabbit) Reagent 3 (ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB High contrast kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. ? Deparaffinise and rehydrate paraffin-embedded tissue sections. ? Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. ? Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. PostBlock (Reagent 2, yellow) 20 min. 8. Washing with wash buffer 3 x 5 min. 9. HRP-polymer (Reagent 3, red) 30 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. We guarantee that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin is used for stabilisation. A material safety data sheet is available upon request.
The Plus HRP Polymer Kit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus HRP Polymer Kit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of horse radish peroxidase (HRP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colorising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus HRP Polymer Kit avoids the problem of background staining caused by endogenous biotin in the tissue
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Endogenous peroxidase activity in the tissue may cause non-specific staining. This enzyme activity can be blocked by incubation with 3% H2O2-solution (peroxide block). Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the HRPpolymer is minimized by incubation with a protein blocking solution (Blocking Solution, provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the enhancement reagent (PostBlock) is applied and incubated. A second washing is followed by the application of the HRP-polymer. Any excess of unbound HRP-polymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the peroxidase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen AEC leads to the formation of a red-brown product of reaction at the place of the target antigen. The chromogen DAB forms a dark brown precipitate.
Reagents provided:
6 ml Blocking Solution Reagent 1 (ready-to-use) 6 ml PostBlock Reagent 2 (ready-to-use) 6 ml HRP-Polymer (Mouse/Rabbit) Reagent 3 (ready-to-use) Substrate systems recommended: Permanent AEC kit, AEC single solution, AEC substrate kit, DAB substrate kit, DAB High contrast kit. Materials required but not supplied: Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O 3% H2O2 solution Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS Pink PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. ? Deparaffinise and rehydrate paraffin-embedded tissue sections. ? Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. ? Tissue sections have to be completely covered with the different reagents in order to avoid drying out.
Procedure:
1. Peroxide blocking (3 % H2O2 solution) 10 min. 2. Washing with wash buffer 1 x 2 min. 3. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 4. Washing with wash buffer 1 x 2 min. 5. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 6. Washing with wash buffer 3 x 5 min. 7. PostBlock (Reagent 2, yellow) 20 min. 8. Washing with wash buffer 3 x 5 min. 9. HRP-polymer (Reagent 3, red) 30 min. 10. Washing with wash buffer 3 x 2 min. 11. AEC or DAB (Controlling the colour intensity via light microscope is recommended.) 5-15 min. 12. Stopping the reaction with distilled H2O when the desired colour intensity is attained 13. Counterstaining and blueing 14. Mounting: aqueous with AEC, permanent with DAB
Expected results:
During the reaction of the substrate with horse radish peroxidase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please read these instructions carefully, contact our technical support . No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pretreatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment. Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. Incubation times were too short or primary antibody concentration too low. 7. Chromogenic substrate solution was too old. Nonspecific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous horse radish peroxidase in the tissue. Maybe the hydrogen peroxide solution used for blocking was inactivated.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous peroxidase or pseudoperoxidase activity may cause non-specific staining. The enzyme activity is blocked by incubation with hydrogen peroxide solution. Tissues containing Hepatitis B Surface Antigen (HBsAg) may give false positive results with HRP (horse radish peroxidase) detection systems (Omata et al, 1980). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the Blocking Solution instead of just draining it away as in other procedures. We guarantee that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin is used for stabilisation. A material safety data sheet is available upon request.
The Plus AP Polymer Kit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Polymer Kit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus AP Polymer Kit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP polymer is minimized by incubation with a protein blocking solution (Blocking Solution, provided with this kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the enhancement reagent (PostBlock) is applied and incubated. A second washing is followed by the application of the AP-polymer. Any excess of unbound APpolymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent AP Red leads to the formation of a magentared product of reaction at the place of the target antigen.
Reagents provided:
100 ml Blocking Solution Reagent 1 (ready-to-use) 100 ml PostBlock Reagent 2 (ready-to-use) 100 ml AP-Polymer (Mouse/Rabbit) Reagent 3 (ready-to-use) Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. PostBlock (Reagent 2, yellow) 20 min. 6. Washing with wash buffer 3 x 5 min. 7. AP-polymer (Reagent 3, red) 30 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent AP Red 10-20 min. (Controlling the colour intensity via light microscope is recommended.) 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: permanent or aqueous with Permanent AP Red
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Therefore, tissues of this origin should be stained with peroxidase detection systems (i.e. POLHRP-125). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the BlockingSolution instead of just draining it away as in other procedures. We will guarantee that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and ProClin 950 used for stabilisation. Material safety data sheets (MSDS) are available upon request.
The Plus AP Polymer Kit is designed for the qualitative detection of antigens in fixed paraffin-embedded tissue sections, in frozen tissue sections, and in cytological samples. It is developed for use in combination with mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. The kit can be used for examining tissues fixed in different solutions, e.g. formalin (neutrally buffered), B5, Bouin, ethanol, or HOPE.
The purpose of the immunohistochemical staining is to make tissue and cell antigens visible. The Plus AP Polymer Kit is a highly sensitive detection kit intended for use in immunohistochemistry and immunocytochemistry. The enzyme polymer in this kit consists of several molecules of secondary antibodies covalently bound to several molecules of alkaline phosphatase (AP). Visualisation occurs via an enzyme-substrate reaction in the presence of a colourising reagent which permits microscopical analysis. The test system is suitable for the detection of mono- and polyclonal primary antibodies and sera obtained from mice and rabbits. In contrast to other detection techniques, which often use the streptavidin-biotin system the Plus AP Polymer Kit avoids the problem of background staining caused by endogenous biotin in the tissue.
Principle of method:
Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP polymer is minimized by incubation with a protein blocking solution (Blocking Solution, provided with this kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the enhancement reagent (PostBlock) is applied and incubated. A second washing is followed by the application of the AP-polymer. Any excess of unbound APpolymer is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Permanent AP Red leads to the formation of a magentared product of reaction at the place of the target antigen.
Reagents provided:
6 ml Blocking Solution Reagent 1 (ready-to-use) 6 ml PostBlock Reagent 2 (ready-to-use) 6 ml AP-Polymer (Mouse/Rabbit) Reagent 3 (ready-to-use) Materials required but not supplied Positive und negative control tissue Xylene or suitable substitutes Ethanol, distilled H2O Reagents for enzyme digestion or heat pre-treatment Wash buffer PBS or TBS PAP Pen Primary antibody (user-defined) Primary antibody diluent Negative control reagent Chromogenic substrate Counter stain solution Mounting medium Cover slips
Storage and handling:
The solutions should be stored at 2-8°C without further dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. They should not be used after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents, please contact our technical support.
Reagent preparation:
Reagents should be at room temperature when used. Deparaffinise and rehydrate paraffin-embedded tissue sections. Pre-treatment (optional) with HIER (Heat Induced Epitope Retrieval) or enzymatic digestion. Tissue sections have to be completely covered with the different reagents in order to avoid drying out
Procedure:
1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 5 min. 5. PostBlock (Reagent 2, yellow) 20 min. 6. Washing with wash buffer 3 x 5 min. 7. AP-polymer (Reagent 3, red) 30 min. 8. Washing with wash buffer 3 x 2 min. 9. Permanent AP Red 10-20 min. (Controlling the colour intensity via light microscope is recommended.) 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: permanent or aqueous with Permanent AP Red
Expected results:
During the reaction of the substrate with alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results which could possibly be caused by the reagents, please contact our technical support. No staining on an actually positive control slide: 1. Reagents were not used in the proper order. 2. Chromogenic substrate solution was too old. 3. Bleaching because chromogen and mounting medium are incompatible. 4. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. Try a pre-treatment such as heat pre-treatment or enzyme digestion. If you used a pre-treatment it should be extended. 5. Primary antibody not from mouse or rabbit, but from a different species. 6. The antigen/epitope was not stable in the fixation and/or pre-treatment procedure used. Try another fixation or pre-treatment Weak staining: 1. Inadequate fixation or overfixation. 2. Incomplete deparaffinisation. 3. The antigen/epitope in the tissue was insufficiently accessible to the primary antibody. If you used heat pre-treatment or enzyme digestion it should be extended. 4. Excessive incubation with Blocking Solution or insufficient washing after this step. 5. Too much wash buffer remains on the slides after washing, diluting the reagents applied in the next step. 6. If you are using PBS-based wash buffer: the activity of alkaline phosphatase in the reagents is blocked if too much wash buffer remains on the slides. 7. Incubation times were too short or primary antibody concentration too low. 8. Chromogenic substrate solution was too old. Non-specific background staining or overstaining: 1. Incomplete deparaffinisation. 2. Excessive tissue adhesive on slides. 3. Insufficient washing especially after the incubation with the enzyme polymer or the chromogenic substrate solution. These washings are critical. 4. Tissue was allowed to (partially) dry out with reagents on. 5. Unspecific binding of the primary antibody. Please use the Blocking Solution provided with this kit or dilute the primary antibody in appropriate diluents. 6. Incubation time of the primary antibody was too long or primary antibody concentration too high. 7. Incubation time of the chromogenic substrate solution was too long or reaction temperature too high (e.g. if temperature in the laboratory is high). 8. The substrate is metabolised by endogenous alkaline phosphatase in the tissue. This undesired activity can often be suppressed using levamisole (see section Limitations of the Procedure).
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Endogenous alkaline phosphatase activity may cause non-specific staining. The enzyme activity can be blocked by incubation with levamisole. However, neither intestinal nor placental alkaline phosphatase can be blocked with levamisole. Therefore, tissues of this origin should be stained with peroxidase detection systems (i.e. POLHRP-125). Inadequate counterstaining and mounting can influence the interpretation of the results. The colour intensity of the reaction product can decrease with time, especially when exposed to light. Overexposure with the protein blocking solution (Blocking Solution) can result in decreasing signal intensity. Therefore, we recommend washing away the BlockingSolution instead of just draining it away as in other procedures. We will guarantee that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid eye, skin or mucous membrane contact with the reagents. In case of a reagent coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagents must be avoided, since otherwise non-specific staining might appear. ProClin 300 and ProClin 950 used for stabilisation. Material safety data sheets (MSDS) are available upon request.
The reactivity of the antiserum is restricted to the Fc part of the IgM molecule. In immunoelectrophoresis and radial immunodiffusion, using various antiserum concentrations against normal human serum a single precipitin line is obtained which shows a reaction of identity with the precipitin line obtained with purified IgM. No precipitation reaction is obtained with purified IgG, IgA, and IgG/Fab fragments. In precipitating techniques as immunoelectrophoresis and radial immunodiffusion to identify the presence of IgM in human serum and other body fluids or to determine its concentration. To prepare an immunoadsorbent for the purification of human IgM from serum or plasma.The antiSerum does not cross-react with any other component of the Human Ig system. Interspecies cross-reactivity is a normal feature of antibodies to immunoglobulins, since Ig of different species frequently share antigenic determinants. In immunoelectrophoresis crossreactivity of this antiSerum has not been observed with Serum of Rhesus Monkey, Cynomolgus and Baboon.
PAX2 is a member of the paired box family of transcription factors, which is required for development and proliferation of the kidney, brain, and mllerian organs. PAX2 genes contain a highly conserved DNA sequence within the paired box region, which encodes a DNA-binding domain, enabling PAX proteins to bind the promoters of specific genes to transcriptionally regulate their expression. PAX2 is specifically expressed in the developing central nervous system, eye, ear, and urogenital tract, and is essential for the development of these organs. In normal adult tissues PAX2 was mainly detected in the urogenital system, including kidney, ureteric epithelium, fallopian tube epithelium, ovary and uterus. In tumors, PAX2 has been detected in renal cell carcinomas, Wilms' tumors, nephrogenic adenomas and papillary serous carcinoma of the ovary. PAX2 has been used as a marker for the identification of renal cell carcinoma and ovarian carcinoma by immunohistochemistry.
Monosan Range:
MONOSAN
Clone:
EP235
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gnarra JR, et al. Cancer Res. 1995; 55:4092-8
References 2:
Mazal PR, et al. Mod Pathol. 2005; 4:535-40
References 3:
Chivukula M, et al. Int J Gynecol Pathol. 2009; 28:570-8
Phosphohistone H3 (PHH3) is a core histone protein, which together with other histones, forms the major protein constituents of the chromatin in eukaryotic cells. In mammalian cells, phosphohistone H3 is negligible during interphase but reaches a maximum for chromatin condensation during mitosis. Immunohistochemical studies showed anti-PHH3 specifically detected the core protein histone H3 only when phosphorylated at serine 10 or serine 28. Studies have also revealed no phosphorylation on the histone H3 during apoptosis. PHH3 can serve as a mitotic marker to separate mitotic figures from apoptotic bodies and karyorrhectic debris, which may be a very useful tool in diagnosis of tumor grades, especially in CNS, skin, gyn., soft tissue, and GIST.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gurley LR, et al. Eur J Biochem 1978; 84:1-15
References 2:
Hendzel MJ, et al. J Biol Chem 1998; 273:24470-8
References 3:
Colman H, et al. Am J Surg Pathol. 2006; 30:657-64
References 4:
Nasr MR, et al. Am J Dermatopathol. 2008; 30:117-22
Alpha-fetoprotein (AFP) is a fetal tumor-associated polypeptide of the albuminoid gene family that binds and transports molecules in addition to many other proposed functions. This secretory protein is synthesized primarily in the fetal liver whereas expression is repressed in adult liver.Anti-AFP has been immunohistochemically demonstrated in hepatocellular carcinoma (HCC) and shows no immunoreactivity in normal liver.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mizejewski GJ et al. Exp Biol Med. 2001; 226:377-408
References 2:
Lazarevich NL et al.Biochemistry (Mosc). 2000; 65:117-33
References 3:
Yusof YA, et al. Anal Quant Cytol Histol. 2003; 25:332-8
Thyroid-stimulating hormone (also known as TSH or thyrotropin) is a peptide hormone synthesized and secreted by thyrotrops in the anterior pituitary gland which regulate the endocrine function of the thyroid gland. TSH is a glycoprotein and consists of two subunits which are non-covalently bound to one another. Anti-TSH reacts with TSH-producing cells (thyrotrophs), and is a useful marker in classification of pituitary tumors.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Batanero E, et al. Brain Behav Immun. 1992; 6:249-64
References 2:
Sanno N, et al. J Clin Endocrinol Metab. 1995; 80:2518-22
References 3:
La Rosa S, et al. Virchows Arch. 2000; 437:264-9
References 4:
Kuzuya N, et al. J Clin Endocrinol Metab. 1990; 71:1103-11
Toxoplasma gondii is a spindle-to-oval-shaped protozoan which presents as an infection in humans of various sorts. The cyst (30 um) and trophozoite (7 um) stages can be identified in humans is such cases. This intracellular parasite is transmitted via raw/undercooked meat, contaminated soil, or by direct contact with an infected host. Infection in humans is usually associated with a variable degree of immunosuppression such as in pregnancy or immunosuppression due to various drugs.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Anti-TdT antibody labels normal cortical thymocytes and primitive lymphocytes. Anti-TdT antibody detects an enzyme found in the nucleus of normal hematopoietic cells, normal cortical thymocytes and in the cytoplasm of megakaryocytes of the bone marrow. TdT expression is seen in over 90% of acute lymphoblastic lymphoma/ leukemia cases with the exception of pre-B-Cell ALL. TdT expression is not seen in normal mature T-or B-lymphocytes. Anti-TdT is positive for approximately one third of all cases of chronic myeloid leukemia, making it a good indicator of better response to chemotherapy.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Motea EA, et al. Biochimica et Biophysica Acta. 2010; 1804:1151-66
References 2:
Stauchen JA, et al. Int J Surg Pathol. 2003; 11:21-4
References 3:
Suzumiya J, et al. J Pathol. 1997; 182:86-91
References 4:
Arber DA, et al. Am J Clin Pathol. 1996; 106:462-8
Anti-Synaptophysin reacts with neuroendocrine cells of human adrenal medulla, carotid body, skin, pituitary, thyroid, lung, pancreas and gastrointestinal mucosa. Positive staining is seen in neurons of the brain, spinal cord, retina, and Paneths cells in the gastrointestinal tract and gastric parietal cells. This antibody identifies normal neuroendocrine cells and neuroendocrine neoplasms. Diffuse, finely granular cytoplasmic staining is observed, which probably correlates with the distribution of the antigen within neurosecretory vesicles. The expression of synaptophysin is independent of the presence of NSE or other neuroendocrine markers. Anti-Synaptophysin is an independent broadrange marker of neural and neuroendocrine differentiation.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Navone F, et al. J Cell Biol. 1986; 103:2511-27
References 2:
Wiedenmann B, et al. Cell. 1985; 41:1017-28
References 3:
Kayser K, et al. Pathol Res Pract. 1988; 183:412-7
Prolactin (PRL) is a single-chain polypeptide of 226 amino acids and plays a role in multiple processes including cell growth, reproduction, and immune function. Anti-Prolactin reacts with prolactin-producing cells and is a useful marker in classification of pituitary tumors and the study of pituitary disease.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Asa SL, et al. Arch Pathol Lab Med. 1982; 106:360-3
References 2:
Duello TM, et al. Am J Anat. 1980; 158:463-9
References 3:
Minniti G, et al. Surg Neurol. 2002; 57:99-103
References 4:
Popadic A, et al. Surg Neurol. 1999; 51:47-54
References 5:
Nevalainen MT, et al. J Clin Invest. 1997; 99:618-27
Protein gene product 9.5 (PGP 9.5), also known as ubiquitin carboxyl-terminal hydrolase-1 (UCH-L1), is a 27-kDa protein originally isolated from whole brain extracts (1). Although PGP9.5 expression in normal tissues was originally felt to be strictly confined to neurons and neuroendocrine cells (2), it has been subsequently documented in distal renal tubular epithelium, spermatogonia, Leydig cells, oocytes, melanocytes, prostatic secretory epithelium, ejaculatory duct cells, epididymis, mammary epithelial cells, Merkel cells, and dermal fibroblasts. LK Campbell et al demonstrated immunostaining of a plethora of different mesenchymal neoplasms with this antibody.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Campbell LK, et al. Mod Pathol. 2003; 16:963-9
References 2:
Bassotti G, et al. J Clin Pathol. 2005; 58:973-7
References 3:
Mahalingam M, et al. J Cutan Pathol. 2001; 28:282-6.
References 4:
Mahalingam M, et al. J Cutan Pathol. 2006; 33:51-6.
Napsin is a pepsin-like aspartic proteinase in the A1 clan of the AA clade of proteinases. There are two closely related napsins, napsin A (NAPSA) and napsin B (NAPSB). Napsin A is involved in processing propeptide pulmonary surfactant protein B (proSP-B) in the lung.4 In normal tissue, Napsin A is expressed in type II pneumocytes of the lung and proximal tubules of the kidney. Napsin A is a useful marker for lung adenocarcinoma
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Brasch F, et al. J Biol Chem. 2003; 278: 49006-14
References 2:
Jagirdar J et al. Arch Pathol Lab Med. 2008; 132:384-96
References 3:
Bishop JA, et al. Hum Pathol. 2010; 41:20-5
References 4:
Ye J, et al. Appl Immunohistochem Mol Morphol. 2011; 19:313-17
References 5:
Mukhopadhyay S, et al. Am J Surg Pathol. 2011; 35:15-25
Immunostaining with anti-myoglobin provides a specific, sensitive, and practical procedure for the identification of tumors of muscle origin. Since myoglobin is found exclusively in skeletal and cardiac muscle and is not present in any other cells of the human body, it may be used to distinguish rhabdomyosarcoma from other soft tissue tumors.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mukai K, et al. Am J Surg Pathol. 1979; 3:373-6
References 2:
Corson JM, et al.Am J Pathol. 1981; 103:384-9
References 3:
Brooks JJ. Cancer. 1982; 50:1757-63
References 4:
Furlong MA, et al. Ann Diagn Pathol. 2001; 5:199-206
Anti-Myeloperoxidase detects granulocytes and monocytes in blood and precursors of granulocytes in the bone marrow. This antibody can detect myeloid cell populations of the bone marrow as well as in other sites.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Pinkus GS, et al. Mod Pathol. 1991; 4:733-41
References 2:
Markoc F, et al. Tumori. 2010; 96:149-53
References 3:
Alexiev BA, et al. Diagn Pathol. 2007; 31;2:42
References 4:
Saravanan L, et al. Int J Lab Hematol. 2010; 32:132-6
References 5:
Manaloor EJ, et al. Am J Clin Pathol. 2000; 113:814-22
Anti-Lysozyme stains myeloid cells, histiocytes, granulocytes, macrophages, and monocytes in human tonsil, colon and skin. It is an important marker that may demonstrate the myeloid or monocytic nature of acute leukemia. The restrictive nature of anti-lysozyme antibody staining suggests that lysozyme may be synthesized predominantly in reactive histiocytes rather than in resting, unstimulated phagocytes. Anti-lysozyme may aid in the identification of histiocytic neoplasias, large lymphocytes and classifying lymphoproliferative disorders.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Rehg J, et al. Toxicol Pathol. 2012; 40: 345-74
References 2:
Seifert RP, et al. Annals Diag Pathol. 2014; 18:253-60.
Luteinizing hormone (LH) is a heterodimeric glycoprotein produced by gonadotropic cells of the pituitary gland. Anti-LH is a useful marker to aid in the classification of pituitary tumors and the study of pituitary disease.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sano T, et al. Virchows Arch A Pathol Anat Histopathol. 1990; 417:361-7
References 2:
Felix I, et al. Hum Pathol. 1991; 22:719-21
References 3:
Saccomanno K, et al. J Clin Endocrinol Metab. 1994; 78:1103-7
Anti-IgM reacts with immunoglobulin mu (IgM) chains. IgM is one of the predominant surface immunoglobulins on B-lymphocytes. This antibody is useful when differentiating and sub-classifying hematolymphoid neoplasms.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Arnold A, et al. New Eng J Med. 1983; 309:1593-1599
References 2:
Leong AS, et al. Geenwich Medical Media Ltd. 1999; 217-219
References 3:
Taylor CR. Arch Path Lab Med. 1978; 102:113-121
References 4:
Kojima M, et al. APMIS. 2002; 110:875-80
References 5:
Pambuccian SE, et al. Am J Surg Pathol. 1997; 21:179-86
Immunoglobulin A (IgA) plays a critical role in mucosal immunity. It is present in the mucosal secretions such as tears, saliva, colostrum, intestinal juice, vaginal fluid, and secretions from the prostate and respiratory epithelium, and represents a key first line of defense against invasion by inhaled and ingested pathogens at the vulnerable mucosal surfaces. It is also found in small amounts in blood. Because it is resistant to degradation by enzymes, secretory IgA can survive in harsh environments such as the digestive and respiratory tracts, to provide protection against microbes that multiply in body secretions. It is useful when identifying multiple myeloma.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ansari NA, et al. Asian Pac J Cancer Prev. 2007; 8:593-6
Human placental lactogen (hPL), also previously known as human chorionic somatomammotropin, is a 22 kD protein with partial homology to growth hormone. hPL is first detectable in the maternal serum in the fifth week of gestation and reaches a plateau by the thirty-fourth week. hPL has been demonstrated by immunochemistry in the syncytiotrophoblastic cells of choriocarcinoma. A rare variant of trophoblastic tumor has been reported in the testis with resemblance to uterine placental site trophoblastic tumor. It consisted purely of intermediate trophoblasts, which was diffusely positive for hPL and focally for ?-hCG.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Shih IM, et al. Am J Surg Pathol. 2004; 28:1177-83
References 2:
Ulbright TM, et al. Am J Surg Pathol. 1997; 21:282-8
Herpes simplex virus is quite ubiquitous and is quite variable in its presentation in human disease. Type I usually infects the non-genital mucosal surfaces. It may affect the skin or internal organs (typically brain, lung, liver, adrenal gland, or GI tract) of immunocompromised individuals. This polyclonal antibody reacts with Type I Herpes viruses. There may be cross-reactivity with varicella zoster virus at higher concentrations. Cross-reactivity with CMV or Epstein-Barr virus is not seen with this antibody.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Helicobacter pylori is strongly associated with inflammation of the stomach and is also implicated in the development of gastric malignancy, peptic ulcers, and gastric lymphomas in humans. Helicobacter pylori can exist in a number of locations: in the mucus, attached to epithelial cells, or inside of vacuoles in epithelial cells, where it produces adhesions that bind to membrane-associated lipids and carbohydrates in or on epithelial cells. The most reliable method for detecting H. pylori infection is a biopsy during endoscopy histologic examination and detection by immunohistochemistry. Immunohistochemical staining of H. pylori on the surface of gastric mucosa is a valuable tool for identification of H. pylori infections.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
hCG is a protein secreted in large quantities by normal trophoblasts; the antibody detects cells and tumors of trophoblastic origin such as Choriocarcinoma. Large Cell Carcinoma and Adenocarcinoma of Lung demonstrate hCG positivity in 90% and 60% of cases respectively. 20% of Squamous Cell Lung Carcinomas are positive for hCG. hCG expression by nontrophoblastic tumors may indicate aggressive behavior since it has been observed that hCG may play a role in the host response to a given tumor.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Granzymes are serine proteases which are stored in specialized lytic granules of cytotoxic T lymphocytes and in natural killer cells. Anti-Granzyme B has been useful in diagnosing Natural killer/T cell lymphoma, as well as anaplastic large cell lymphoma. High percentages of cytotoxic T cells have been shown to be an unfavorable prognostic indicator in Hodgkins disease.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kummer JA, et al. Clin Exp Immunol. 1995; 100:164-72
Glucose transporter type I (GLUT1), a prototype member of GLUT super family, reacts with a 55 kD protein, is a membrane-associated erythrocyte glucose transport protein. It is a major glucose transporter in the mammalian blood-brain barrier, and also mediates glucose transport in endothelial cells of the vasculature, adipose tissue and cardiac muscle. GLUT1 is detectable in many human tissues including those of colon, lung, stomach, esophagus, and breast. GLUT1 is overexpressed in malignant cells and in a variety of tumors that include the breast, pancreas, cervix, endometrium, lung, mesothelium, colon, bladder, thyroid, bone, soft tissues, and oral cavity. Immuohistochemical detection of GLUT1 can discriminate between reactive mesothelium and malignant mesothelioma. Anti-GLUT1 with anti-Claudin1, and anti-EMA are perineurial markers in diagnosis of perineuriomas. Anti-GLUT1 is also useful in distinguishing benign endometrial hyperplasia from atypical endometrial hyperplasia and adenocarcinoma. GLUT1 expression has been associated with increased malignant potential, invasiveness, and a poor prognosis in general. Expression of GLUT1 is a late event in colorectal cancer and expression in a high proportion of cancer cells is associated with a high incidence of lymph node metastases.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kato Y, et al. Mod Pathol. 2006; 20:215-20
References 2:
Afify A, et al. Acta Cytol. 2005; 49:621-6
References 3:
Parente P, et al. J Exp Clin Cancer Res. 2008; 27:34
Anti-Glucagon antibody detects glucagon-secreting cells and tumors such as glucagonomas. Studies show that approximately 80% of glucagonomas are malignant and these patients have a syndrome often initially recognized by dermatologists. Symptoms include necrolytic migratory erythema as well as diabetes, anemia, stomatitis, weight loss, frequent venous thromboses, and in some instances, diarrhea and psychiatric disturbances. The diagnosis may be readily confirmed by the demonstration of elevated plasma glucagon concentration.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Quesada I, et al. J Endocrinol. 2008; 199:5-19
References 2:
Gurlo T, et al. J Histotechnol. 2016; 39:8-16
References 3:
Wewer Albrechtsen NJ, et al. Biomark Med. 2016; 10:1141-51
Anti-GH is a useful marker in classification of pituitary tumors and the study of pituitary disease (acromegaly). It reacts with GH-producing cells. Growth hormone receptors have been found in various non-pituitary cells, including that from hepatocellular carcinoma and various benign and malignant cutaneous lesions.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Rezaei M, et al. J Res Med Sci. 2012; 17:681-5
References 2:
Al-Brahim NY, et al. J Clin Pathol. 2006; 59:1245-53
References 3:
Fukaya T, et al. Cancer. 1980; 45:1598-1603
References 4:
Kovacs K, et al. Virch Arch Pathol Anat. 1982; 395:59-68
Anti-Gastrin antibody gives positive staining of G-cells of human antral/pyloric mucosa and cells producing gastrin or a structural gastrin analogue as is seen in stomach; no staining of other cells or tissue types has been observed. This antibody may react with sulfated and non-sulfated forms of gastrin. The antibody cross-reacts with more than 50% of the present choleocystokinin octapeptide.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kasacka W, et al. Folia Morphol. 2012; 71:39-44.
References 2:
Hur K, et al. J Cancer Res Clin Oncol. 2006; 132:85-91
References 3:
Waldum et al. Frontiers in Endocrinology. 2017; 8:1-7
Anti-FSH is a useful marker in classification of pituitary tumors and the study of pituitary disease. It reacts with FSH-producing cells (gonadotrophs).
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Baenziger JU, et al. Biochim Biophys Acta. 1988; 947:287-306
References 2:
Nussey SS, et al. BIOS Scientific Publishers Ltd; 2001 p. 217-79
References 3:
Uccella S, et al. Pituitary. 2000; 3:131-9
References 4:
Schmid M, et al. Pathol Res Pract. 2001; 197:663-9
Anti-Factor VIII-Related Antigen antibody reacts with endothelial cells and neoplastic blood cells. This antibody has helped to establish the endothelial nature of some lesions of disputed histogenesis, e.g. Kaposis sarcoma and cardiac myxoma. Not all endothelial cells synthesize (or store) this molecule; therefore, it should not be surprising that not all tumors of endothelial differentiation (benign or malignant) react with this antigen.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Nichols GE, et al. Am J Clin Pathol. 1992; 97:770-5
References 2:
Falk S, et al. Am J Surg Pathol. 1993; 17:959-70
References 3:
Meis-Kindblom JM, et al. Am J Surg Pathol. 1998; 22:683-97
References 4:
Allison KH, et al. Am J Surg Pathol. 2004; 28:298-307
References 5:
Peyvandi F, et al. Blood Transfus. 2011; 9 Suppl 2:s3-8
The claudins are a family of over twenty proteins which are components of tight junction. Tight junctions are specialized regions of cell to cell contact; made up of network of strands to act as a molecular gasket for preventing the leakage of ions, water etc. between cells. They are abundant in luminal epithelial sheets where they maintain epithelial cell polarity. The claudins constitute a variable component, with specific claudins being associated with specific tissues. The immunoreactivity for anti-claudin 1 is membranous and is found in nearly all carcinomas. The staining is much stronger in the carcinoma cells than in normal tissues. Anti-claudin 1 in a panel of immunostains with EMA (positive), S-100 (negative), and GLUT1 can be utilized as a robust marker in diagnosis of perineurioma and neurofibroma. Recently studies showed anti-claudin seems to be a specific marker for meningiomas. Therefore, anti-claudin with anti-EMA, anti-S-100 protein, anti-CD34, and anti-glial fibrillary acidic protein may be helpful in the diagnosis of meningiomas from histologic mimics
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Folpe AL, et al. Am J Surg Pathol. 2002; 26:1620-6
Anti-CEA specifies a group of proteins in the Carcinoembryonic Antigen (CEA) family of proteins which are present in the epithelia of various types and tumors (both benign and malignant) derived from such epithelia. Such tissues are represented by the epithelia of colon, bronchus, alveoli, breast, pancreas, biliary tract, superficial layer and parietal layers of the stomach. Predominately biliary canaliculi are labelled in the liver and this factor is useful in the diagnosis of hepatocelluar carcinoma. Anti-CEA has been quite useful in differentiating adenocarcinoma of the lung vs. mesothelioma. Associated products: CK 5/6, Calretinin, WT-1, E-Cadherin, TTF-1, TAG-72, EMA, CK 20
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Shield PW, et al. Am J Clin Pathol. 1996; 105:157-62
References 2:
Sheahan K, et al. Am J Clin Pathol. 1990; 94:157-64
Anti-CD3 antibody has been considered the best all around T-cell marker. This antibody reacts with an antigen present in early thymocytes. The positive staining of this marker may represent a sign of early commitment to the T-cell lineage.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Beverley PC, et al. Eur J Immunol. 1981; 11:329-34
References 2:
Clevers H, et al. Eur J Immunol. 1988; 18:705-10
References 3:
Hedvat CV, et al. Hum Pathol. 2002; 33:968-74
References 4:
Karube K, et al. Am J Surg Pathol. 2003; 27:1366-74
Immunohistochemical staining with anti-calcitonin antibody has proven to be an effective way of demonstrating calcitonin-producing cells in the thyroid. C-cell hyperplasia and medullary thyroid carcinomas stain positive for calcitonin. Studies of calcitonin have resulted in the identification of a wide spectrum of C-cell proliferative abnormalities.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Matias-Guiu X, et al. Endocr Pathol. 2014; 25:21-9
References 2:
Fisher S, et al. Arch Pathol Lab Med. 2008;132:359-72
Complement component C3 plays a central role in the activation of complement system. Its activation is required for both classical and alternative complement activation pathways. C3d deposition in the renal transplant PTCs (peritubular capillaries) is indicative of AR (acute rejection) with subsequent high probability of graft loss. Anti-C3d, combined with anti-C4d, can be utilized as a tool for diagnosis of AR and warrant prompt and aggressive anti-rejection treatment. In another study, Pfaltz et al. have shown that anti-C3d labeled the epidermal basement membrane in 97% (31/32) cases of bullous pemphigoid (BP), with none of the normal controls demonstrating such findings. In the same study 27% (3/11) cases of pemphigus vulgaris (PV) demonstrated intercellular C3d deposition. Therefore, C3d immunohistochemistry is a helpful adjunct in the diagnosis of BP (and perhaps PV), especially in the cases in which only formalin-fixed, paraffin embedded tissue is available for analysis.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Bickerstaff A, et al. Am J Pathol. 2008; 173:347-57
References 2:
Kuypers DR, et al. Transplantation. 2003; 76:102-8
ACTH or Adrenocorticotropic hormone is synthesized from pre-pro-opiomelanocortin (pre-POMC). ACTH is produced and secreted from corticotrophs in the anterior lobe (or adenohypophysis) of the pituitary gland. The anti-ACTH immunohistochemical reagent could be useful in the study of neoplastic and non-neoplastic pituitary diseases
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Pizarro CB, et al. Braz J Med Biol Res. 2004; 37:235-43
References 2:
Kageyama K, et al. Am J Med Sci. 2002; 324:326-30
References 3:
Fan X, et al. J Histochem Cytochem. 2002; 50:1509- 16
References 4:
Japon MA, et al. J Clin Endocrinol Metab. 2002; 87:1879-84
The immunohistochemical staining of Alpha-1-Antitrypsin is considered to be very useful in the study of inherited AAT deficiency, benign and malignant hepatic tumors and yolk sac carcinomas. Positive staining for A-1-Antitrypsin may also be used in detection of benign and malignant lesions of an histiocytic nature. Sensitivity and specificity of the results have made this antibody a useful tool in the screening of patients with cryptogenic cirrhosis or other forms of liver disease with portal fibrosis of uncertain etiology.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Callea F, et al. J Hepatol. 1986; 2:389-401
References 2:
Palmer PE, et al.Am J Clin Pathol. 1974; 62:350-4
References 3:
Palmer PE, et al. Cancer. 1980; 45:1424-31
References 4:
Raintoft I, et al. Hum Pathol. 1979; 10:419-24
References 5:
Ramsay AD, et al. Appl Immunohistochem Mol Morphol. 2008; 16:140-7
Rabbit anti Human caspase-7 antibody recognizes an epitope within the C-terminal region (CT) of Caspase-7, a ~35 kDa cysteine protease, otherwise known as ICE-like Apoptotic Protease 3 (ICE-LAP3).
Caspase-7, a member of the ICE/Ced-3 subfamily, is an executioner caspase which undergoes proteolytic cleavage of its precursor to form active p12 and p20 subunits. Evidence that activation of Caspase-7 occurs during cell death induced by the cytokine death receptors Fas/APO-1 and the receptor of tumour necrosis factor (TNFR-1), coupled with the fact that granzyme-B activated Caspase-7 cleaves the nuclear enzyme poly (ADP-ribose) polymerase (PARP), suggests an important role for Caspase-7 in both cytokine-mediated and granzyme-B mediated apoptosis pathways.
Rabbit anti caspase-4 (N-terminal) antibody recognizes an epitope within the N-terminal region (NT) of Caspase-4, otherwise known as ICH-2. Caspase-4, a member of the Caspase-1 subfamily of cysteine proteases, exists as an inactive pro-enzyme which undergoes proteolytic cleavage of its p30 precursor form, into smaller p20 and p10 subunits.The cellular localization of Caspase-4 on the endoplasmic reticulum (ER) membrane, has resulted in this protein becoming a focus of studies in which dysfunction or stress to the ER membrane is implicated, such as Alzheimers disease and Ischemia and confirms the involvement of Caspase-4 as an instigator of cellular apoptosis.
Rabbit anti Human Caspase-1 antibody recognizes an epitope within the C-terminal region (CT) of human Caspase-1, otherwise known as IL-1Beta converting enzyme. Caspase-1 is an intracellular cysteine protease, identified as a mammalian homologue to C. elegans cell death gene (ced-3).Caspase-1 has been classified as an inflammatory, rather than apoptotic caspase, due to its essential role in the cleavage of the inactive precursors of the cytokines IL-1beta and IL-18, into their mature activated and secretable forms. Regulation of pro-inflammatory cytokines by Caspase-1 has made inhibitors of Caspase-1 a possible target for use as therapeutic drugs for the treatment of inflammatory diseases (Ghayur et al. 1997).Rabbit anti Human Caspase-1 antibody detects a cleaved subunit band of approximately 21 kDa in human heart cell lysates (predicted precursor MWT 45.2kDa).
Rabbit anti Mycobacterium tuberculosis polyclonal antibody recognizes PPD from Mycobacterium tuberculosis. Rabbit anti M. tuberculosis has not been cross absorbed and may react with related micro-organisms, however the antibody is non-reactive with E.coli K12, Salmonella typhimurium, Pseudomonas aeruginosa, Streptococcus (group B), Candida albicans and Neisseria meningitidis.
Lysozyme is a 14 kd enzyme directed against the b 1 a 4 glycosidic bond between N-acetylglucosamine and N-acetylmuramic acid residues that make up peptidoglycan. Lysozyme is an antimicrobial protein secreted by polymorphonuclear leukocytes and is widely distributed in secretions such as airway secretions and nasal fluid whereas it is the most effective antimicrobial protein. It is also produced by monocytes, macrophages and epithelial cells. Lysozyme is able to kill bacteria by enzymatic lysis of bacterial cell walls and by a nonenzymatic mechanism. Allthough lysozyme is highly active against many gram-positive bacteria it is ineffective against gram-negative bacteria unless potentiated by certain cofactors (lactoferrin, antibody-complement or hydrogen peroxide-ascorbic acid). Next to its antimicrobial activity lysozyme has many other physiological functions including inactivation of certain viruses, important roles in surveillance of membranes of mammalian cells, immune regulatory activity, anti-inflammatory and antitumor activity
Human lactoferrin (LF) is an 80 kDa glycoprotein which was first isolated from human milk. It plays an important part in the immune system and helps to fight infections. Lactoferrin promotes the health of the gastro-intestinal system by improving the intestinal microbial balance. In addition, LF can be found in epithelia and most body fluids and secretions. Lactoferrin is secreted in plasma by neutrophils. Its plasma concentration also represents a positive relation to the total pool of neutrophils and the rate of neutrophil turnover. In inflammation lactoferrin is released from secondary granules of neutrophilic leukocytes into the extracellular medium. Therefore the extracellular lactoferrin concentration can be used as an index for neutrophil activation. Lactoferrin strongly binds to iron and this iron binding property is considered to be an important antimicrobial. Human lactoferrin binds to bacterial products through its highly positively charged N-terminus, it kills various bacteria, most probably by inducing intracellular changes in these bacteria without affecting the membrane permeability. Cleavage by pepsin of lactoferrin leads to the release of lactoferricin H. This 47 amino acid peptide has more antimicrobial activity than its precursor and it can inhibit the classical but not the alternative complement pathway. Lactoferrin also plays a role in signal transduction, immunomodulation and has antiadhesive, anticancer, antiviral activity.
Lactoferrin is an approximately 80 kDa glycoprotein which was first isolated from milk and found in epithelia and most body fluids and secretions. Lactoferrin is secreted in plasma by neutrophils. Its plasma concentration represents a positive relation to the total pool of neutrophils and the rate of neutrophil turnover. In inflammation lactoferrin is released from secondary granules of neutrophilic leukocytes into the extracellular medium. Therefore the extracellular lactoferrin concentration can be used as an index for neutrophil activation. <br /> Lactoferrin is able to strongly bind to iron and considered to have antibacterial properties. Human lactoferrin binds to bacterial products through its highly positively charged N terminus and kills various bacteria most probably by inducing intracellular changes in these bacteria without affecting the membrane permeability. Lactoferrin also plays a role in signal transduction, immunomodulation and has antiadhesive, anticancer, antiviral activity.
Sheep anti Human C3c antibody recognizes the C3c component of human complement, formed as a result of the inactivation of C3b. Sheep anti Human C3c antibody may be used for the detection of C3 deposits in tissues following complement activation.
SLP65 / BLNK (SH2 domain-containing leukocyte-specific phosphoprotein of 65 kDa; B cell linker protein), also known as BASH, is an adaptor protein that plays key role in B cell activation initiated by cross-linking the B cell receptor (BCR). Phosphorylated by Syk tyrosine kinase, SLP65 serves as a scaffold for Btk tyrosine kinase, Vav1 guanine nucleotide exchange factor, phospholipase C gamma2, as well as Grb2 and Nck adaptor proteins; thus represents a central linker protein that bridges the BCR-associated kinases with a multitude of signaling pathways.
Monosan Range:
MONOSAN
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
PAG (phosphoprotein associated with GEMs), also known as Cbp (Csk-binding protein), is a ubiquitously expressed 46 kDa transmembrane adaptor protein present in membrane rafts (glycosphingolipid-enriched microdomains), which however migrates on SDS PAGE gels anomalously as an 80 kDa molecule. Following tyrosine phosphorylation by Src family kinases, PAG binds and thereby activates the protein tyrosine kinase Csk, the major negative regulator of the Src family kinases. Signaling via the B-cell receptor in B cells or high affinity IgE receptor (FcepsilonRI) in mast cells leads to PAG increased tyrosine phosphorylation and Csk binding, while T cell receptor signaling causes PAG dephosphorylation, loss of Csk binding and increased activation of the protein tyrosine kinase Lck.
Monosan Range:
MONOSAN
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
SLP76 (SH2 domain-containing leukocyte protein of 76 kDa) is a cytosolic adaptor protein which translocates to the plasma mambrane and is involved in multiple signaling pathways in T cells, mast cells, neutrophils and platelets; B cells express its analog SLP65/BLNK (B cell linker protein). SLP76 is phosphorylated by Syk-family and Tec-family tyrosine kinases and couples them to the phosphorylation and activation of PLC-gamma. Via Gads or Grb2, SLP76 also associates with LAT adaptor by involvement of SLP76 proline-rich region. The SH2 domain of SLP76 has been identified as the region involved in binding the serine/threonine kinase HPK1. HPK1 may act as both a positive and a negative regulator by promoting the Jnk-mitogen activated protein kinase (MAPK) pathway and inhibiting the pathway leading to AP-1 activation.
Monosan Range:
MONOSAN
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
LIME (Lck-interacting molecule) is a 30 kDa double-palmitoylated protein with unusually basic cytoplasmic domain, expressed by T cells. After ligation of CD4 or CD8 T cell coreceptors, LIME is phosphorylated by Src-family kinases and associates with Lck and Fyn kinases and with their negative regulator Csk. Interestingly, Csk-mediated phosphorylation of C-terminal negative-regulatory tyrosine of LIME-associated Lck can result in increase of enzymatic activity compared with the total pool of Lck, thus, LIME serves as a positive regulator of TCR-dependent T cell signaling. However, under some circumstances, LIME may mediate inhibitory signals.
Monosan Range:
MONOSAN
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The antiserum against the vesicular acetylcholine transporter is a unique immunohistochemical marker for cholinergic nerves, more specific than the commonly used acetylcholinesterase (AchE), since it does not react with postsynaptic neurons, and is more sensitive than choline acetyltransferase (ChAT). The antiserum recognizes VAChT both in the CNS and PNS. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: frozen sections of human small intestine (Stefanini fixation).
Located in the cell membrane of thyroid cells, NIS can allow sodium and iodine flow across the membrane. The antiserum is raised against the C-terminus of rat NIS. Suitable for studying the sensitivity of thyroid tumors to iodine treatment. Absorption with 50-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: frozen sections of rat thyroid.
Useful for studying sensory afferent neurons. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: frozen sections of rat spinal cord.
The antiserum is raised against a synthetic peptide (SHMSTSAPPP) from the C-terminus of the rat CCK-A receptor. Suitable for labelling the receptors for the gastrointestinal hormone and neuropeptide CCK. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: frozen sections of rat pancreas.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 100 µl dist. water
Storage:
2-8°C
References 1:
Ohlsson, B. et al. J. Gastroenterol. 2000;35: 612-8
The antibody reacts with a large variety of tumors with oncofetal characteristics; the antigen may also be detected in normal epithelia and tissue of non-neoplastic state. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: Bouin-fixed paraffin sections of human colon carcinoma.
Histidine decarboxylase (HDC) is the enzyme catalyzing the conversion of histidine into histamine. HDC can be found in the histamine secreting ECL cells of some species as well as in the mast cells. Absorption with 10-100 ug immunogen per ml diluted antiserum abolishesthe staining. <br>In Western blot the antiserum detects the 54 kDa and 73 kDa forms in addition to a 63 kDa form (rat stomach, see Dartsch et al., 1998).<br>Positive control: Stefanini-fixed frozen sections of rat fundus. <br> Dartsch, C., Chen, D. & Persson, L. Multiple forms of rat stomach histidine decarboxylase may reflect posttranslational activation of the enzyme. Regul. Pept. 77, 33-41 (1998).
Ornithine decarboxylase is the rate-limiting enzyme in polyamine biosynthesis converting ornithine into putrescine. In normal tissue ornithine decarboxylase activity is low but increases in proliferating tissue. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: Stefanini-fixed frozen sections of renal cortex from testosterone-treated mice.
Nitric oxide synthase is an enzyme catalizing the synthesis of NO from L-arginine. The antiserum was raised against a synthetic peptide of rat cerebellar NOS, that shows no homology with other related proteins. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining. Positive control: frozen sections of rat colon.
Phenylethanolamine-N-methyltransferase (PNMT) is an enzyme converting noradrenaline to adrenaline. <br>The enzyme is present in adrenomedullary cells and in the brain neurons. <br> Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining.<br>Positive control: DEPC-fixed paraffin sections of rat adrenal gland.
Adrenomedullin is a potent vasorelaxing and hypotensive peptide, originally isolated from human pheochromocytoma. Adrenomedullin shares some homology with CGRP (calcitonin gene-related peptide). The antiserum was raised using synthetic peptides as immunogens. The antibody does not cross-react with CGRP. Positive control: Stefanini-fixed frozen sections of rat fundus.
The CGRP-sequence was predicted from the corresponding mRNA. CGRP is present in the C-cells of the thyroid and in nerves, in both brain and periphery, particularly within the sensory system. CGRP is a potent vasodilator, probably involved in neurogenic inflammation. Medullary carcinomas of the thyroid contain large amounts of CGRP. Absorption with 10-100 ug CGRP per ml diluted antiserum abolishes the staining while calcitonin does not. Does not cross-react with amylin (IAPP).<br>Positive control: frozen sections of rat colon.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 100 µl dist. water
Storage:
2-8°C
References 1:
Schulze, E. et al. Acta Histochem.1997; 99: 301309
References 2:
Bechakra, M. et al. Mol.Pain.2018; 14, 1744806918797040
The intestinal peptide YY is related to the PP-family of peptides and occurs in the glicentin cells in the gut. They are numerous in the rectum, colon, and ileum and few in the duodenum and jejunum. PYY has hormone-like action, inhibits gut motility and pancreatic exocrine secretion and cause vasoconstriction. <br>PYY may occur in endorine tumors of the pancreas and of the rectum. Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining.<br>Positive control: frozen sections of rat intestine.
The CGRP-sequence was predicted from the corresponding mRNA. CGRP is present in the C-cells of the thyroid and in nerves, in both brain and periphery, particularly within the sensory system. CGRP is a potent vasodilator, probably involved in neurogenic inflammation. Medullary carcinomas of the thyroid contain large amounts of CGRP. Absorption with 10-100 ug CGRP per ml diluted antiserum abolishes the staining while calcitonin, does not. Cross-reacts with amylin (IAPP).<br>Positive control: Stefanini-fixed frozen sections of rat colon.
Substance P occurs in nerve fibers of the central and peripheral nervous system and in endocrine cells of the gut. It stimulates smooth muscle contraction, gives rise to vasodilation and is involved in sensory functions. Substance P-containing tumors arising in the ileum are often associated with the carcinoid syndrome, characterized by flushing of the skin, diarrhea, broncho-constriction and sudden drops in blood pressure. Substance P is commonly found in the midgut carcinoids and some of the symptoms may be related to this peptide. Absorption with 10-100 ug SP and NKA per ml diluted antiserum abolishes the staining while GRP and NKB do not. Positive control:<strong> </strong>frozen sections of rat colon.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 100 µl dist. water
Storage:
2-8°C
References 1:
Kressel, M.et al. J. Comp. Neurol. 1999;412: 161172
Gastrin-secreting cells are numerous in the antrum and a few are found in the proximal duodenum. The antibody can be used for the diagnosis of gastrin-producing tumors which are mainly found in the pancreas and occasionally in the stomach and the duodenum. <br>Absorption with 10-100 ug gastrin 1-34 and CCK 8 per ml antiserum abolishes the staining. Positive control: formalin-fixed paraffin sections of rat antrum.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 100 µl dist. water
Storage:
2-8°C
References 1:
Portela-Gomes, G. M.et al. Histochem. Cell Biol. 1999;111: 4954
References 2:
Portela-Gomes, G. M.et al. J. Histochem. Cytochem.1997;45: 81522
References 3:
Mulder, H. et al. Gastroenterology 1994;107: 7129
VIP is localized in nerve fibers of the central and peripheral nervous system, and is probably acting as a neurotransmitter. Smooth muscle relaxation, vasodilation and secretion from exocrine glands are some of the effects of VIP. The Verner-Morrison or Watery Diarrhea Hypokaliemia and Achlorhydria (WDHA) syndrome is a characteristic clinical syndrome associated with overproduction of VIP from endocrine tumors. These VIP-producing tumors are usually neuroblastomas of endocrine tumors in the pancreas.<br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining, while PHI does not.<br>Positive control: Stefanini-fixed frozen sections of rat intestine.
Glucagon is a common constituent of endocrine pancreatic tumors and of rectal carcinoids. The antibody is specific to pancreatic glucagon. Absorption with 10-100 ug glucagon per ml diluted antiserum abolishes the staining. Positive control: Bouin-fixed paraffin sections of cat pancreas.
The vesicular monoamine transporter is responsible for the vesicular uptake of monoamines, like dopamine, norepinephrine, epinephrine, serotonin and histamine. The antiserum recognizes monoaminergic neurons of the CNS, the ECL-cells of the stomach, as well as enteric nerve fibers. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. In Western blot experiments using a crude synaptic vesicle fraction of rat brain, the 70 kDa of anti-VMaT2 is recognized (suggested dilution 1:500). Positive control: frozen sections of rat small intestine.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 100 µl dist. water
Storage:
2-8°C
References 1:
Carballo-Carbajal, I. et al. Nat.Commun. 2019;10: 973
Ornithine decarboxylase is the rate-limiting enzyme in polyamine biosynthesis converting ornithine into putrescine. In normal tissue ornithine decarboxylase activity is low but increases in proliferating tissue. Positive control: Stefanini-fixed frozen sections of renal cortex from testosterone-treated mice.
Nitric oxide synthase is an enzyme catalizing the synthesis of NO from L-arginine. The antiserum was raised against a synthetic peptide of rat cerebellar NOS, that shows no homology with other related proteins. <br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining. Positive control: frozen sections of rat colon.
Histamine is a neurotransmitter in the central nervous system as well as a mast cell constituent. In addition, histamine is produced by endocrine cells (ECL-cells) in the oxynthic mucosa of the stomach. Absorption with 10-100 ug histamine per ml diluted antiserum abolishes the staining, while noradrenaline, 5-HT, VIP, glucagon and histidine do not. Positive control: cryostat sections of carbodiimide fixed human skin or freeze-dried paraffin-sections (vapor fixed in diethylpyrocarbonate; DEPC) of rat stomach.
Pancreastatin is a fragment of chromogranin A and is produced by proteolytic processing of chromogranin A in several peptide hormone-producing cells, such as pancreatic islet cells and gut endocrine cells, as well as tumors arising from these cells. <br> Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining. Positive control: Bouin-fixed paraffin sections of rat pancreas.
Insulin is produced by the B-cells of the pancreatic islets and by insulin-producing islet cell tumors. <br>Absorption with 10-100 ug proinsulin per ml diluted antiserum inactivates the antiserum, while C-peptide and insulin only partly reduce staining.<br>Positive control: formalin-fixed paraffin sections of human pancreas.
Monosan Range:
MONOSAN
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 100 µl dist. water
Storage:
2-8°C
References 1:
Kohnert, K. D. et al. Regul. Pept.1999; 82: 719
References 2:
Ahrén, B. et al. Pancreas 1999; 18: 7583
References 3:
Al-Amily, I. et al. Pflugers.Arch.2019; 471:633-645
The peptide was found in the salivary gland venom of the lizard Gila monster. It belongs to the VIP-secretin family of peptides and has many VIP-like actions. Helospectin-immunoreactivity has been demonstrated in the neurons of the gut.<br> Absorption with 10-100 ug helospectin per ml diluted antiserum abolishes the staining, while secretin, PHI and PACAP do not. <br> Cross-reacts with helodermin. Positive control: Stefanini-fixed frozen sections of rat small intestine (1% carbodiimide added to the fixative).
Regulates the level of GABA available for secretion via secretory vesicles. Marker for neurons using GABA as signaling substance. <br> Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining. Positive control: frozen sections of rat cerebellum.
GAP-43, also called neuromodulin, B-50, pp46 and F1, is a neuron-specific, membrane-associated phosphoprotein involved in axonal growth and found in neurons undergoing regeneration.<br>The antiserum was raised against a synthetic C-terminal peptide of GAP-43.<br> Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes staining. Positive control: rat duodenum.
Pituitary adenylate cyclase activating peptide (PACAP), originally isolated from ovine hypothalamus, belongs to the VIP-family of peptides. PACAP-related peptide (PRP) is a 29-amino acid peptide, which is produced together with PACAP-27 and PACAP-38 when PreproPACAP is being processed. PRP is abundant in the brain, but can be also found in the respiratory and gastrointestinal tracts.<br>Absorption with 10-100 ug immunogen per ml diluted antiserum abolishes the staining, while PACAP-27, PACAP-38, VIP, PHI, CRF, oxytocin and vasopressin do not.<br>Positive control: Stefanini-fixed frozen sections of rat duodenum, fundus or antrum.
Serotonin is produced by endocrine cells of the stomach, duodenum and ileum. The polyclonal antibody to serotonin can be used to differentiate tumors of serotoninergic origin. The antigen localization is cytoplasmic. <br>Absorption with 10-100 ug serotonin per ml diluted antiserum abolishes the staining. Positive control: duodenum.
Neurokinin A/Substance K represents a member of the tachykinin family of peptides. Neurokinin A, which arises by cleavage of the substance P precursor, occurs in neurons in the central and peripheral nervous systems, and is particularly numerous in the gastrointestinal tract. The biological actions of neurokinin A are similar to those of substance P, and include vasodilation and stimulation of smooth muscle contraction. <br>Absorption with 10-100 ug NKA per ml diluted antiserum abolishes the staining, while substance P does not. Positive control: formalin-fixed paraffin sections of rat colon.
Neuropeptide Y is a peptide belonging to the PP-family and occurs in neurons and adrenal medullary cells. The brain contains large quantities of NPY and it is also found in the peripheral nervous system, where it coexists with noradrenaline in sympathetic fibers. NPY inhibits gut motility and causes vasoconstriction. Pheochromocytomas contain NPY. Absorption with 10-100 ug NPY 1-20 per ml diluted antiserum abolishes the staining, while PYY and PP do not. Positive control: Stefanini-fixed sections of rat small intestine.
Oct-2 is a transcription factor of the POU homeo-domain family that binds to the Ig gene octamer sites, regulating B-cell-specific genes. These are involved in proliferation and differentiation and, despite the scarce evidence for Oct-2 expression in T cells, it has been shown that this factor participates in transcriptional regulation during T-cell activation. Oct-2 activity is dependent on phosphorylation and alternatitive splicing.Various lymphomas are also positive for this marker including the following: Bchronic lymphocytic leukemia, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, plasmacytoma, Burkitt lymphoma, diffuse large cell lymphoma, diffuse large B-cell lymphoma, T-cell rich B-cell lymphoma, nodular lymphocyte predominant Hodgkin lymphoma, and classic Hodgkin lymphoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Browne P, et al. Am J Clin Pathol. 2003; 120:767-77
References 2:
García-Cosío M, et al. Mod Pathol. 2004; 17:1531-8
References 3:
Gibson SE, et al. Am J Clin Pathol. 2006; 126:916-24
Anti-Cytokeratin (OSCAR) is well suited to distinguish epithelial carcino¬ma from non-epithelial malignancies and is used to aid epithelial tumor classification. This antibody has been used to characterize the source of various neoplasms and to study the distribution of keratin containing cells in epithelia during normal development and during the develop¬ment of epithelial neoplasms. This antibody stains cytokeratins present in normal and abnormal human tissues and has shown high sensitivity in recognizing epithelial cells, and carcinomas.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
OSCAR
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gown, AM, et al. Am J Clin Pathol 1985;84:413
References 2:
Battifora, H. Am J Surg Pathol 1988;12:24
References 3:
Lewis JE et al. Hum Pathol. 1997 Jun;28(6):664-73
References 4:
Mueller JD et al. Cancer. 2000 Nov 1;89(9):1874-82
Mi is a transcription factor implicated in pigmentation, bone development and in mast cells. Various forms of Mi exist ranging from 50-70 kD in size. This antibody targets the 52-56 kD range. This antibody has been useful in identifying malignant melanoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
C5/D5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Liegl B, et al. Am J Surg Pathol. 2008; 32:608-14
References 2:
Righi A, et al. Int J Surg Pathol. 2008; 16:16-20
References 3:
Weinreb I, et al.. Virchows Arch. 2007; 450:463-70
References 4:
Ohsie SJ, et al. J Cutan Pathol. 2008; 35:433-44
References 5:
Hornick JL, et al. Am J Surg Pathol. 2008; 32:493-501
GCDFP-15 is a glycoprotein localized in the apocrine metaplastic epithelium lining breast cysts and in apocrine glands in the axilla, vulva, eyelid, ear canal, and in salivary glands. GCDFP-15 positivity is seen in breast carcinomas. On the other hand, colorectal carcinomas, lung carcinoma, mesotheliomas rarely stain with this antibody. Because of its specificity for breast carcinoma, this antibody is often helpful in distinguishing metastasis of unknown primary.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP1582Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mazoujian G, et al. Am J Pathol. 1983; 110:105-12
References 2:
Liegl B, et al. Histopathology. 2007; 50:439-47
References 3:
Bhargava R, et al. Am J Clin Pathol. 2007; 127:103-13
References 4:
Tornos C, et al. Am J Surg Pathol. 2005; 29:1482-9
References 5:
Takeda Y, et al. Arch Pathol Lab Med. 2008; 132:239-43
Factor XIIIa has been identified in platelets, megakaryocytes, and fibroblast-like mesenchymal or histiocytic cells in the placenta, uterus, and prostate, monocytes and macrophages and dermal dendritic cells. Anti- Factor XIIIa has been found to be useful in differentiating between dermatofibroma (almost all cases +), dermatofibrosarcoma protuberans (-/+) and desmoplastic malignant melanoma. Anti-factor XIIIa positivity is also seen in capillary hemagioblastoma, hemangioendothelioma, hemangiopericytoma, xanthogranuloma, xanthoma, hepatocellular carcinoma, glomus tumor, and meningioma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP3372
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Nemes Z, Hum Pathol 1992 Jul; 23(7):805-10
References 2:
Horenstein MG et al. Am J Surg Pathol. 2000 Jul;24(7):996-1003
References 3:
Kraus MD et al. Am J Dermatopathol. 2001 Apr;23(2):104-11
References 4:
Abenoza P, et al. Am J Dermatopathol. 1993; 15:429-34
References 5:
Anstey A, Cerio R, et al.: Am J Dermatopathol 1994 Feb: 16(1):14-22
Blood group Lewis carbohydrate determinants are oligosaccharides on glycolipids and glycoproteins. In healthy individuals the LewisY antigen is a type 2 antigen usually only expressed in low levels of a few cell types such as some epithelial cells. Reportedly these antigens are aberrantly expressed in high levels in many carcinomas. Anti-BG8, LewisY reactivity in immunohistochemistry is seen in carcinomas of the breast, lung, and colon
Antibody Isotype:
IgM
Monosan Range:
MONOSAN Ready To Use
Clone:
F3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Davidson B, et al. Virchows Arch. 1999; 435:43-9
References 2:
King JE, et al. Histopathology. 2006; 48:223-32
References 3:
Marchevsky AM et al. Appl Immunohistochem Mol Morphol. 2007; 15:140-4
CD99, as detected with a variety of antibodies, is expressed by virtually almost all Ewings sarcoma and primitive peripheral neuroectodermal tumors (ES/PNET) and demonstrates strong and diffuse membranous staining. Other tumors that may show CD99 expression include neuroendocrine carcinomas, mesenchymal chondrosarcomas, solitary fibrous tumors, synovial sarcomas, vascular tumors, small round blue cell tumors, lymphoblastic lymphoma, acute myeloid leukemia, and myeloid sarcoma.5 However, strong and diffuse membranous reactivity for CD99 favors ES/PNET over the other diagnostic considerations. The other CD99+ tumors usually show cytoplasmic and more heterogeneous staining. Therefore, when making a final diagnostic interpretation, CD99 must be considered in a panel with other antibodies.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EPR3097Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Rettig WJ, et al. Lab Invest. 1992; 66:133
References 2:
Fellinger EJ, et al. Amer J Surg Pathol. 1992; 16:746
References 3:
Ambros IM, et al. Cancer. 1991; 139:317
References 4:
Khoury JD. Adv Anat Pathol. 2005; 12:212-20
References 5:
Dabbs DJ. Theranostic and Genomic Applications. 2014; 126
CD71, also known as transferrin receptor, is a membrane glycoprotein that mediates the uptake of iron from transferrin for hemoglobin synthesis in erythroid cells. Early erythroid precursors and erythroblasts contain the highest mass of transferrin receptors, and expression is lost as these cells cease hemoglobin synthesis and mature into erythrocytes. Therefore, anti-CD71 is a useful marker for highlighting erythroid precursors in bone marrow specimens. Increased CD71 expression is also associated with active cell growth including neoplastic tumor growth and may be seen in various carcinomas such as thyroid carcinomas, lung carcinomas, breast carcinomas, hepatocellular carcinomas and colorectal carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-48
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ponka P, et al. Int J Biochem Cell Biol. 1999; 31:1111-1137
References 2:
Sieff C, et al. Blood. 1982; 60:703-713
References 3:
Lesley J, et al. Cell Immunol.1984; 83:14-25
References 4:
Nakahata T, et al. Leuk Lymphoma. 1994; 13:401-409
References 5:
Marsee DK, et al. Am J Clin Pathology. 2010; 13:429-435
CD23 antigen is a 45-60 kDa membrane glycoprotein identified as a low affinity receptor for IgE production as well as a receptor for lymphocyte growth factor. CD23 is found in some mature B-cell lymphomas and in Reed-Sternberg cells in Hodgkin disease. Follicular dendritic cells and some activated B-cells within germinal centers express CD23 in high density and mantle zone B-cells are stained weakly. The majority of chronic lymphocytic leukemias/small lymphocytic lymphomas are CD23 positive, whereas mantle cell lymphomas are generally negative, so this marker is useful when applied with other markers to separate the small cell lymphomas. Precursor B and T lymphomas, myeloid neoplasms, and mature T-cell lymphomas are CD23 negative and other small cell lymphomas are occasionally positive. CD23 is also positive on activated mature B-cells expressing IgM or IgD, monocytes/ macrophages, follicular dendritic cells, T-cell subsets, eosinophils, Langerhans cells and small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL).
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-57
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
CD21 (also known as complement receptor 2 (CR2), C3d receptor, or EBV receptor) is a 140 kDa membrane protein on B-lymphocytes to which the Epstein-Barr virus (EBV) binds during infection of these cells. The antigen is absent on T-lymphocytes, monocytes, and granulocytes. MON 3028 is useful in the identification of follicular dendritic cell matrix found in normal lymph node and tonsillar tissue. This antibody also labels follicular dendritic cell sarcomas. Anti-CD21 is valuable in differentiating follicular lymphoma with marginal zone differentiation from marginal zone lymphoma with follicular involvement. It also plays a role in distinguishing among nodular lymphocyte predominant Hodgkin lymphoma, lymphocyte-rich classic Hodgkin lymphoma, and T-cell/histiocyte-rich B-cell lymphoma in combination with other B-cell and T-cell markers. Anti-CD21 is also useful in identifying abnormal follicular dendritic cell pattern in angioimmunoblastic T-cell lymphoma and follicular T-cell lymphoma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP3093
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Dillon KM et al. J Clin Pathol. 2002 Oct;55(10):791-4
CD2 is a surface antigen present on all peripheral T-cell lymphocytes. CD2 is associated with signaling and mediating adhesion to other T-cells through the lymphocyte function-associated antigen and CD48/BCM1 complex. MON 3225 is a useful immunohistochemical reagent for identification of normal T-lymphocytes, and for assessing lymphoid malignancies.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-11
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Aguilera NS, et al. Arch Pathol Lab Med. 2006; 130:1772-9
References 2:
Barrionuevo C, et al. Appl Immunohistochem Mol Morphol. 2007; 15:38-44
References 3:
Bovenschen HJ, et al. Br J Dermatol. 2005; 153:72-8
References 4:
Foon KA, et al. Blood. 1986; 68:1-31
References 5:
Gonzalez L, et al. Journal of Comparative Pathology 2001; 125:41-7
Smoothelin is a constituent of the smooth muscle cell cytoskeleton protein exclusively found in differentiated smooth muscle cells (SMC). Cells with SMC-like characteristics, such as myofibroblasts and myoepithelial cells, as well as skeletal and cardiac muscle do not contain smoothelin.1,2 To distinguish bladder muscularis mucosae (MM) from muscularis propria (MP) muscle bundles is crucial for accurate staging of bladder carcinoma. Strong smoothelin expression is nearly exclusively observed in muscularis propria. Therefore, the staining pattern of MP (strongly positive) and MM (negative or weakly positive) makes this technique an attractive diagnostic tool for the sometimes difficult task of staging bladder urothelial carcinoma, such as in transurethral resection specimens of urinary bladder tumors.3-5 Differentiating between smooth muscle tumors and other mesenchymal neoplasms of the GI tract can be challenging in small biopsies. Anti-smoothelin immunostaining can be helpful in differentiating benign (+) from malignant smooth muscle tumors (-), and other mimics(-).
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
R4A
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
van der Loop FT, et al. J Cell Biol. 1996; 134:401-11
References 2:
Maake C, et al. J Urol. 2006; 175:1152-7
References 3:
Paner GP, et al. Am J Surg Pathol. 2010; 34:792-9
References 4:
Council L, et al. Mod Pathol. 2009; 22:639-50
References 5:
Coco DP, et al. Am J Surg Pathol. 2009; 33:1795-801
T-cell leukemia/lymphoma protein 1 (TCL1, TCL1A, p14TCL1) is a 14 kDa product of the TCL1 gene that is involved in T-cell prolymphocytic leukemia (T-PLL). TCL1 protein is normally found in the nucleus and cytoplasm of lymphoid lineage cells during early embryogenesis. TCL1 is expressed in differentiated Bcells under both reactive and neoplastic conditions, antigen committed B-cells, and in germinal center B-cells. The Anti-TCL1 immunohistochemical reactivity is reportedly useful identifying B-cell lymphomas including follicular lymphoma and Burkitt lymphoma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-7
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Takizawa J, et al. Jpn J Cancer Res. 1998; 89:712-8
References 2:
Narducci MG, et al. Cancer Res. 2000; 60:2095-100
References 3:
Rodig SJ, et al. Am J Surg Pathol. 2008; 32:113-22
References 4:
Herling M, et al. Leukemia. 2006; 20:280-5
References 5:
Pescarmona E, et al. Histopathology. 2006; 49:343-8
S100P is a member of the S100 family of proteins. The family is expressed in a wide range of cells and is thought to play a role in cell cycle progression and in differentiation. Anti-S100P with nuclear or nuclear/cytoplasmic immunoreactivity can be seen in pancreatic ductal adenocarcinomas, while it is rarely detectable in benign pancreatic ducts. It may also help to distinguish urothelial carcinomas from other genitourinary neoplasms such as prostate carcinoma
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
16/f5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Lin F, et al. Am J Surg Pathol 2008; 32:78-91
References 2:
Deng HB. Am J Clin Pathol 2008; 129:81-8
References 3:
Nakata K et al. Hum Pathol 2010; 41:824-31
References 4:
Higgins JP, et al. Am J Surg Pathol 2007; 31:673-80
CD33, also known as gp67 or SIGLEC-3, is a 67 kDa glycosylated transmembrane protein that is a member of the sialic acid-binding immunoglobulin-like lectin (siglec) family. Although the precise physiological function of CD33 is unknown, it may mediate cell to cell adhesion and modulate inflammatory and immune response. In normal tissue, anti-CD33 labels myeloid cells (especially myeloid precursors), liver Kupffer cells, lung alveolar macrophages, and placental syncytiotrophoblasts. In neoplastic tissue, anti-CD33 is useful for the identification of acute myeloid leukemia.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN Ready To Use
Clone:
PWS44
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Freeman SD, et al. Blood. 1995; 85:2005-12
References 2:
Laszlo GS, et al. Oncotarget. 2016; 7:43281-94
References 3:
Crocker, PR et al. Biochem Soc Symp 2002; 69:83-96
CD11c is an adhesion receptor of the leukocyte function-associated family of molecules. Reportedly CD11c is expressed in hairy cell leukemia whereas the majority of other small B-cell lymphomas do not express CD11c antigen. This indicates that immunohistochemical staining of formalin-fixed biopsies with anti-CD11c can be useful for identification of hairy cell leukemia.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
5D11
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Korinna, J et al. Pathobiology 2008; 75:252256
References 2:
Jones, G et al. Br J Hemaetol 2011; 156:186-195
References 3:
Went, PT et al. Am J Surg Pathol 2005; 29:474478
References 4:
Miranda, RN et al. Modern Pathology 2000; 13:13081314
Neuron-specific enolase (NSE) is the glycolytic isoenzyme of the enolase gamma-gamma dimer specifically detected in neurons of neuroendocrine cells, and their corresponding tumors. In addition, NSE has been demonstrated immunohistochemically in the non-neoplastic cells of the pituitary, peptide secreting tissues, pineolocytes, neuroendocrine cells of the lung, thyroid, parafollicular cells, adrenal medulla, islets of Langerhans, Merkel cells of the skin, and melanocytes. Anti-NSE immunostaining is also positive in normal striated muscle, hepatocytes and, to a lesser extent, smooth muscle. Anti-NSE is a useful marker to identify peripheral nerves.5 When used for the identification of neuroendocrine differentiation, it is suggested that it be employed in a panel with more specific markers such as anti-synaptophysin, anti-chromogranin, and anti-neurofilament.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-55
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Wick MR, et al. Am J Clin Pathol. 1983; 79:703-7
References 2:
Vinores SA, et al. Arch Pathol Lab Med. 1984; 108:536-40
Human germinal center associated lymphoma (HGAL) protein is specifically expressed in the cytoplasm of germinal center B-cells, but is absent in mantle and marginal zone B-cells and in the interfollicular and paracortical regions in normal tonsils and lymph nodes. Its high degree of specificity for germinal center B-cells makes anti-HGAL an ideal marker for the detection of germinal center-derived B-cell lymphomas. Anti-HGAL has the highest overall sensitivity of detecting follicular lymphoma (FL) and in detecting the interfollicular and diffuse components of FL compared with antibodies against bcl2, LMO2, CD10, and bcl6. The addition of anti-HGAL to the immunohistologic panel is beneficial in the work-up of nodal and extranodal B-cell lymphomas, and the efficacy of anti-HGAL in detecting the follicular, interfollicular, and diffuse components of FL is of particular value in the setting of variant immunoarchitectural patterns
Antibody Isotype:
IgG2a-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-49
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Natkunam Y, et al. Blood. 2005; 105:397986
References 2:
Natkunam Y, et al. Blood. 2007; 109:298-305
References 3:
Younes SF, et al. Am J Surg Pathol. 2010; 34:1266-76
References 4:
Higgins RA, et al. Arch Pathol Lab Med. 2008; 132:441-6
Phosphohistone H3 (PHH3) is a core histone protein, which together with other histones, forms the major protein constituents of the chromatin in eukaryotic cells. In mammalian cells, phosphohistone H3 is negligible during interphase but reaches a maximum for chromatin condensation during mitosis. Immunohistochemical studies showed anti-PHH3 specifically detected the core protein histone H3 only when phosphorylated at serine 10 or serine 28. Studies have also revealed no phosphorylation on the histone H3 during apoptosis. PHH3 can serve as a mitotic marker to separate mitotic figures from apoptotic bodies and karyorrhectic debris, which may be a very useful tool in diagnosis of tumor grades, especially in CNS, skin, gyn., soft tissue, and GIST.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gurley LR, et al. Eur J Biochem 1978; 84:1-15
References 2:
Hendzel MJ, et al. J Biol Chem 1998; 273:24470-8
References 3:
Colman H, et al. Am J Surg Pathol. 2006; 30:657-64
References 4:
Nasr MR, et al. Am J Dermatopathol. 2008; 30:117-22
T-bet, a T-box transcription factor, is expressed in CD4+ T-lymphocytes committed to T-helper (Th)1 Tcell development from naïve T-helper precursor cells (Thp) and redirects Th2 T-cells to Th1 development. Anti-T-bet is a marker of mature T-cells and is expressed at very low levels in Thp cells and is absent in precursor T-lymphoblastic leukemia/lymphoma cells. Scattered small lymphocytes in the interfollicular T-cell zone of reactive lymphoid tissue, including tonsil, lymph node, and spleen exhibited nuclear staining for anti-T-bet, with no anti-T-bet staining observed in germinal centers or mantle or marginal zones. T-bet is expressed in a significant subset of B-cell lymphoproliferative disorders, particularly at an early stage of B-cell development (precursor B-cell lymphoblastic leukemia/lymphoblastic lymphoma), and B-cell neoplasms derived from mature B-cells, including CLL/SLL, marginal zone lymphoma, and hairy cell leukemia.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-46
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Szabo SJ, et al. Cell. 2000; 100:665-69
References 2:
Jöhrens K, et al. Am J Surg Pathol. 2007; 31:1181-5
References 3:
Atayar C, et al. Am J Pathol. 2005; 166:127-34
References 4:
Dorfman DM, et al. Am J Clin Pathol. 2004; 122:292-7
IgG4-related sclerosing disease has been recognized as a systemic disease entity characterized by an elevated serum IgG4 level, sclerosing fibrosis, and diffuse lymphoplasmacytic infiltration with the presence of many IgG4-positive plasma cells. Clinical manifestations are apparent in the pancreas, bile duct, gall bladder, lacrimal gland, salivary gland, retroperitoneum, kidney, lung, breast, thyroid, and prostate. Immunohistochemical analyses in the case of IgG4-related sclerosing disease not only exhibit significantly more than normal IgG4-positive plasma cells in affected tissues but also significantly higher IgG4/IgG ratios (typically > 30%).
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-44
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sakata N, et al., Am J Surg Pathol. 2008; 32:553-9
References 2:
Dhobale S, et al., J Clin Rheumatol. 2009; 15:354-7
References 3:
Li Y, et al., Pathol Int. 2009; 59:636-41
References 4:
Koyabu M et al., J Gastroenterol. 2010; 45:732-41
References 5:
Kamisawa T, et al., World J Gastroenterol. 2009; 21:2357-60
CD56, also known as neural cell adhesion molecule (NCAM), is a calcium-independent homophilic binding protein that belongs to a group of cell adhesion molecules including cadherins, selectins, and integrins. CD56 is involved in cellcell adhesion of neural cells during embryogenesis and is expressed on most neuroectodermally derived tissues.1-3 In normal tissue, anti-CD56 labels neurons, glia, schwann cells, NK (natural killer) cells, and a subset of T-cells.3 CD56 expression can be seen in most NK cell neoplasms, certain subtypes of T-cell lymphoma and in some plasma cell neoplasms.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-42
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Langdon, SP et al. Cancer Research 1988; 48(21):6161-6165
References 2:
Kaufmann, O et al. Hum Pathol. 1997 Dec; 28(12): 1373-8
References 3:
Tao, J et al. Am J Surg Pathol. 2002 Jan; 26(1):111-8
References 4:
Ely, SA et al. Am J Pathol. 2002 Apr; 160(4): 1293-9
References 5:
Sumi, M et al. Leuk Lymphoma. 2003 Jan; 44(1): 201-4
CD14 is a 55kDa glycosyl-phosphatidylinositol-linked membrane protein, involved in endotoxin binding and recognition of apoptotic cells. CD14 is expressed on monocytes, macrophages, follicular dendritic cells, and granulocytes. Anti-CD14 can detect these cells, including monocyte-derived cells which are frequently increased in diffuse large B-cell lymphoma (DLBCL), as well as in neoplastic cells in acute myeloid leukemia with monocytic differentiation and chronic myelomonocytic leukemia.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EPR3653
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gregory CD, et al. Apoptosis. 1999; 4:11-20
References 2:
Wright SD, et al. Science. 1990; 249: 1431-33
References 3:
Marmey B, et al. Hum Pathol. 2006; 37:68-77
References 4:
Smeltzer JP, et al. Clin Cancer Res. 2014; 20:2862-72
References 5:
Rollins-Raval MA, et al. Histopathology. 2012; 60: 933-42
p120 catenin is encoded on chromosome 11q11. Alpha-catenin and beta-catenin bind to the intracellular domain of E-cadherin while p120 catenin binds E-cadherin at a juxta-membrane site. The complex stabilizes tight junctions. In the cell, p120 catenin localized to the E-cadherin/catenins cell adhesion complex, directly associates with cytoplasmic C-terminus of E-cadherin and may similarly interact with other cadherins. A deficiency of E-cadherin results in the intracytoplasmic accumulation of p120 catenin. Lobular carcinoma of the breast shows intracytoplasmic accumulation of p120 catenin while ductal carcinoma shows reduced membrane p120 catenin without cytoplasmic accumulation. In gastric and colonic carcinoma, strong cytoplasmic p120 catenin is associated with discohesive infiltrative morphology.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Reynolds AB, et al. Oncogene.; 7:2439-45 (1992)
References 2:
Thoreson MA, et al. J Cell Biol.; 148:189-202 (2000)
References 3:
Sarrio D, et al. Oncogene.; 23:3272-83 (2004)
References 4:
Dabbs DJ, et al. Am J Surg Pathol.; 31:427-37 (2007)
The antibody s useful as an immunohistochemical reagent to stain melanocytes and tumors derived therefrom. Anti-PNL2 reactivity is identified in the cytoplasm of cutaneous and oral mucosal melanocytes. Anti-PNL2 labels intraepidermal nevi, while the dermal components of compound nevi are largely non-reactive.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
PNL2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Complement component C3 plays a central role in the activation of complement system. Its activation is required for both classical and alternative complement activation pathways. C3d deposition in the renal transplant PTCs (peritubular capillaries) is indicative of AR (acute rejection) with subsequent high probability of graft loss. Anti-C3d, combined with anti-C4d, can be utilized as a tool for diagnosis of AR and warrant prompt and aggressive anti-rejection treatment. In another study, Pfaltz et al. have shown that anti-C3d labeled the epidermal basement membrane in 97% (31/32) cases of bullous pemphigoid (BP), with none of the normal controls demonstrating such findings. In the same study 27% (3/11) cases of pemphigus vulgaris (PV) demonstrated intercellular C3d deposition. Therefore, C3d immunohistochemistry is a helpful adjunct in the diagnosis of BP (and perhaps PV), especially in the cases in which only formalin-fixed, paraffin embedded tissue is available for analysis.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Bickerstaff A, et al. Am J Pathol. 2008; 173:347-57
References 2:
Kuypers DR, et al. Transplantation. 2003; 76:102-8
The antibody is a monoclonal, anti-melanoma antibody that reacts with an antigen that has yet to be identified.1 Notably used as a melanoma marker, KBA.62 also detects smooth muscle, basal cells of the epidermis and hair shaft epithelia of the skin.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
KBA.62
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Pages C, et al.. Hum Pathol. 2008; 39:1136-42
References 2:
Aung P, et al. Am J Surg Pathol. 2012; 36:265-72
References 3:
E Cohen-Knafo et al. J Clin Pathol. 1995; 48:826-831
References 4:
Kaufmann O, et al.. Mod Pathol, 1998 Aug; 11(8):740-6
CD3s immunohistochemical detection locates the cytoplasmic component of CD3 protein. Anti-CD3 is considered to be a pan-T-cell marker and reacts with an antigen present at the surface and in the cytoplasm of T lymphocytes. Anti-CD3 is widely used for the identification of immature and mature T-cell malignancies.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-39
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Beverley PC, et al. Eur J Immunol.; 11:329-34 (1981)
References 2:
Hedvat CV, et al. Hum Pathol.; 33:968-74 (2002)
References 3:
Karube K, et al. Am J Surg Pathol.; 27:1366-74 (2003)
References 4:
Dogan A, et al. Am J Surg Pathol.; 27:903-11 (2003)
Varicella Zoster Virus (VZV), a member of the human herpes virus family, causes two distinct clinical manifestations: chickenpox and shingles. Primary VZV infection results in chickenpox (varicella), which may rarely result in complications including encephalitis or pneumonia. Even when clinical symptoms of chickenpox have resolved, VZV remains dormant in the nervous system of the infected person (virus latency), in the trigeminal and dorsal root ganglia. In about 10-20% of cases, VZV reactivates later in life producing a disease known as herpes zoster or shingles. Serious complications of shingles include postherpetic neuralgia, zoster multiplex, myelitis, herpes ophthalmicus, or zoster sine herpete. VZV is closely related to the herpes simplex virus (HSV). Affected skin shares so many histological similarities that distinguishing between them may be difficult. Immunohistochemistry with anti-VZV appears quite sensitive and specific on formalin-fixed paraffin-embedded tissues in the distinction between HSV and VZV.
Antibody Isotype:
N/A
Monosan Range:
MONOSAN Ready To Use
Clone:
SG1-1, SG1-SG4, NCP-1 & IE-62 (7 clone cocktail)
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kleinschmidt D, et al. J Neurol Sci. 1998 Aug 14; 159(2):213-8
References 2:
Kaye SB, et al. Br J Ophthalmol. 2000 Jun;84(6):563-71
References 3:
A.F. Nikkels, et al. Virchows Archiv A pathol Anat. 1993; 422:121-126
Thyroglobulin (Tg) is the precursor of the iodinated thyroid hormones thyroxine (T4) and triiodothyronine (T3). Tg is a high molecular weight glycoprotein found in normal thyroid follicular cells. Thyroglobulin is useful for identifying thyroid carcinoma of papillary and follicular types and for identifying tumors of thyroid origin when working with adenocarcinoma of unknown primary
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-41
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sellitti DF and Suzuki K. Thyroid. 2014; 24:625-38
References 2:
Bellet D, et al. J Clin Endocrinol Metab 1983; 56:530-3
References 3:
Bejarano PA, et al. Appl Immunohistochem Mol Morphol. 2000; 8:189-94
PAX-8 is a transcription factor expressed during embryonic development of Müllerian organs, kidney, and thyroid, with continued expression in some epithelial cell types of these mature tissues.1 It can be useful for marking several types of carcinoma including ovarian serous carcinoma, clear cell renal cell carcinoma, and papillary thyroid carcinoma.1-5 Additionally, PAX-8 is not found in the epithelial cells of the breast, lung, mesothelium, stomach, colon, pancreas and other sites.1-4
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ozcan A, et al. Mod Pathol. 2011; 24:751-64
References 2:
Laury AR, et al. Am J Surg Pathol. 2011; 35:816-26
References 3:
Nonaka, D et al. Am J Surg Pathol. 2008; 32:1566-71
Kidney-specific cadherin (Ksp-cadherin) is a member of the cadherin family of cell adhesion molecules that is found exclusively in the basolateral membrane of renal tubular epithelial cells of the distal tubules and collecting duct. Ksp-cadherin may be useful in distinguishing between renal neoplasms of distal nephron origin from those of proximal tubule origin.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-33
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Hemoglobin alpha chain belongs to the globin family and is involved in oxygen transport from the lung to the various peripheral tissues. Hemoglobin A is comprised of two alpha chains and two beta chains. Immunohistochemical localization of hemoglobin is excellent as an erythroid marker for the detection of immature, dysplastic, and megaloblastic erythroid cells in myeloproliferative disorders, such as erythroleukemia. In contrast, myeloid cells, lymphoid cells, plasma cells, histiocytes, and megakaryocytes do not stain with anti-hemoglobin A.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EPR3608
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
OMalley DP, et al. Mod Pathol. 2005; 18:1550-61
References 2:
Cherie H Dunphy, et al. Appl Immun Mol Morphol, 2005; 15(2):154-159
Anti-GCDFP-15, mouse monoclonal (23A3), and anti-mammaglobin, mouse monoclonal (304-1A5) and rabbit monoclonal (31A5), is an antibody cocktail. GCDFP-15 is a 15 kDa glycoprotein which is localized in the apocrine metaplastic epithelium lining breast cysts and in apocrine glands in the axilla, vulva, eyelid, and ear canal. Mammaglobin (10 kDa) is a breast-associated glycoprotein distantly related to the secretoglobin family that includes human uteroglobin and lipophilin.This antibody cocktail is useful in identifying breast carcinoma.
Antibody Isotype:
IgG2a/IgG1/IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
23A3+(304-1A5+31A5)
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Cohen, C et al. Arch Pathol Lab Med. 1993; 117:291-4
References 2:
Bhargava R, et al. Am J Clin Pathol. 2007; 127:103-13
References 3:
Takeda Y, et al. Arch Pathol Lab Med. 2008; 132:239-43
References 4:
Tornos C, et al. Am J Surg Pathol. 2005; 29:1482-9
PU.1 is a transcription factor that has been shown to be important for normal B-cell development. PU.1 belongs to the ETS family of transcription factors. It is expressed in the myeloid lineage and in immature as well as mature B-lymphocytes, with the exception of plasma cells. PU.1 is essential during early B-cell differentiation. The absence of PU.1 results in total block of B-cell development at the prepro stage. Very little is known about PU.1 function in later stages of B-cell development. PU.1 does not seem to play a role in the end-stage of B-cell development and is not expressed in plasma cells. PU.1 exerts an important role in the regulation of the expression of crucial B-cell proteins, such as immunoglobulin (Ig) genes, and CD20 and its putative binding sites were also identified in the promoters of CD79, CD10, and CD22. PU.1 binds to the 3 enhancer region of both the Ig kappa and lambda light chain genes and it also regulates the immunoglobulin heavy chain genes through the intron enhancer region.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EPR3158Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
The parathyroid glands function within the endocrine system to promote blood calcium homeostasis through controlled release of parathyroid hormone (PTH). This process involves the synthesis and secretion of PTH by activated parathyroid chief cells during conditions of hypocalcemia. With the anatomical proximity to the thyroid and capacity of associated neoplasms of the parathyroid to mimic thyroid tumors, challenges can arise in distinguishing between these types of abnormalities. In cases where there is uncertainty about a tumor being of parathyroid origin, immunohistochemical evaluation using anti-PTH can be of value.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-31
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Aldinger KA, et al. Cancer; 49:388-97 (1982)
References 2:
Brown EM. Mineral Electrolyte Metal; 8:130-50 (1982)
References 3:
Abate EG, et al. Front Endocrinol (Lausanne).; 7:172 (2017)
References 4:
Duan K, et al. Turk Patoloji Derg.; 31 Suppl 1:80-97 (2015)
References 5:
Chen HL, et al. Journal of Biology and Chemistry; 277:19374-81 (2002)
p21 is one of the inhibitors of the phosphorylation of the cyclin-cdk complex. p21, which is an inhibitor of G1 cdks, suppresses the cell cycle and inhibits DNA synthesis. Although p21 is induced by p53 and inhibits cdk (cyclin-dependent kinase) activity, there was virtually no correlation between the expression of p21 and that of p53; this finding was consistent with two reports, though another reported an inverse correlation between the expression of p21 and that of p53. p53independent expression of p21 might account for the discrepancy between the expression of p53 and that of p21. It is expressed in normal human tissue and a wide array of tumors.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
DCS-60.2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Herpes simplex virus is quite ubiquitous and is quite variable in its presentation in human disease. Type I usually infects the non-genital mucosal surfaces. It may affect the skin or internal organs (typically brain, lung, liver, adrenal gland, or GI tract) of immunocompromised individuals. This polyclonal antibody reacts with Type I Herpes viruses. There may be cross-reactivity with varicella zoster virus at higher concentrations. Cross-reactivity with CMV or Epstein-Barr virus is not seen with this antibody.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Granzymes are serine proteases which are stored in specialized lytic granules of cytotoxic T lymphocytes and in natural killer cells. Anti-Granzyme B has been useful in diagnosing Natural killer/T cell lymphoma, as well as anaplastic large cell lymphoma. High percentages of cytotoxic T cells have been shown to be an unfavorable prognostic indicator in Hodgkins disease.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kummer JA, et al. Clin Exp Immunol. 1995; 100:164-72
CDX-2 is a caudal-related homeobox transcription factor whose expression in the adult is normally present in the gastrointestinal (GI) epithelium. It is implicated in the development and maintenance of the intestinal mucosa. This protein is expressed immunohistochemically in the nuclei of normal GI epithelium. CDX-2 protein expression has been seen in GI carcinomas. Anti-CDX-2 has been useful to establish GI origin of metastatic adenocarcinomas and carcinoids and is especially useful to distinguish metastatic colorectal adenocarcinoma from lung adenocarcinoma. However, mucinous carcinomas of the ovary also stain positively with this antibody, which limits the usefulness of this marker in the distinction of metastatic colorectal adenocarcinoma versus mucinous carcinoma of the ovary.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EPR2764Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mazziotta RM, et al. Appl Immunohistochem Mol Morphol. 2005; 13:55-60
References 2:
Erickson LA, et al. Endocr Pathol. 2004; 15:247-52
References 3:
Saqi A, et al. Am J Clin Pathol. 2005; 123:394-404
References 4:
Saad RS, et al. Am J Clin Pathol. 2004; 122:421-7
References 5:
Kaimaktchiev V, et al. Mod Pathol. 2004; 17:1392-9
MART-1 (also known as Melan A) is a melanocyte differentiation antigen. MART-1 is a transmembrane protein present in melanocytes of normal skin, retina, nevi, and most melanomas. MART-1 is a very useful marker for identifying metastatic melanomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
A103
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kageshita T et al. J Immunother 1997 Nov;20(6):460-5
References 2:
Yaziji H, et al. In J Surg Pathol. 2003 Jan;11(1):11-5
References 3:
Mocellin S et al. J Immunother. 2001 Nov-Dec;24(6):447-58
References 4:
Perez RP et al. Hum Pathol. 2000 Nov;31(11):1381-8
References 5:
Hoang MP et al. J Cutan Pathol. 2001 Sep;28(8):400-6
Cytokeratin 5 is an intermediate filament protein of 58 kD amongst the cytokeratin family. It is a type II (basic) cytokeratin. Antibodies to this protein identify basal cells of squamous and glandular epithelia, myoepithelia, and mesothelium.1 Cytokeratin 14 is a 50 kD polypeptide found in basal cells of squamous epithelia, some glandular epithelia, myoepithelium, and mesothelial cells.1 Anti-cytokeratin 5 has been useful in the differential diagnosis of metastatic carcinoma in the pleura versus epithelial mesothelioma.2 Anti-cytokeratin 14 has been demonstrated to be useful in differentiating squamous cell carcinomas from other epithelial tumors.3,4 Anti-Cytokeratin 5, along with anti-cytokeratin 14, has been found to have an application in identifying the basal-like phenotype of breast carcinoma.5
Antibody Isotype:
IgG /IgG3
Monosan Range:
MONOSAN Ready To Use
Clone:
EP1601Y & LL002
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Dabbs DJ. Elsevier Saunders, 2014. Print. P. 212
References 2:
Comin CE, et al. Am J Surg Pathol. 2007; 31:1139-48
References 3:
Reis-Filho JS, et al. Appl Immunohistochem Mol Morphol. 2003; 11:1-8
The antibody labels a 50kDa, multiple myeloma oncogen-1 (MUM1) protein. MUM1 is encoded by the MUM1/IRF-4 gene, which is mapped to 6q23-25 and identified as a myeloma-associated oncogene. It is a member of the interferon regulatory factor family of transcription factors and plays an important role in the regulation of gene expression in response to signaling by interferon and other cytokines. MUM1 positive cells express the protein in the nucleus in a diffuse and microgranular pattern. However, some positivity is also observed in the cytoplasm of MUM1-expressing cells. In normal/reactive lymphoid tissues, such as lymph node, this antibody stains plasma cells, some B-cells in the light zone of germinal centers, and a subset of T-cells (T-cells in germinal centers and interfollicular areas). MUM1 expression has been described in diffuse large B-cell lymphoma (DLBCL). MON 3318 can stain other Bcell lymphomas such as lymphoplasmacytic lymphoma, chronic lymphocytic leukemia, follicular lymphoma, marginal zone lymphoma, lymphoblastic lymphoma/leukemia, primary effusion lymphoma, DLBCL, Burkitt-like lymphoma, and classical Hodgkin lymphoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-8
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Falini B, et al. Blood. 2000; 95:2084-92
References 2:
Grossman A, et al. Genomics. 1996; 37:229-33
References 3:
Neresh KN. Haematologica. 2007; 92:267-8
References 4:
Van Imhoff GW, et al. J Clin Oncol. 2006; 24:4135-42
References 5:
Gualco G, et al. Appl Immunohistochem Mol Morphol. 2010; 18:301-10
Mammaglobin is breast-associated glycoprotein distantly related to secretoglobin family that includes human uteroglobin and lipophilin. Anti-mammaglobin labels cytoplasm of normal breast epithelial cells as well as primary and metastatic breast carcinomas. Absence of mammaglobin expression is typically seen in prostate, kidney, colon, rectum, small intestine, stomach, pancreas, lung and thyroid tissue.2,5 Mammaglobin may be used as part of an immunohistochemical panel for determination of metastatic breast carcinoma and tumor of unknown primary origin.
Antibody Isotype:
IgG1, IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
304-1A5/31A5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Fleming TP et al. Ann NY Acad Sci 2000;923:78-89
References 2:
Bhargava R, et al. Am J Clin Pathol. 2007; 127:103-13
References 3:
Wang Z, et al. Int J Clin Exp Pathol. 2009; 2:384-9
References 4:
Han JH, et al. Arch Pathol Lab Med. 2003; 127:1330-4
C4d is a stable split product remnant of classical complement activation which becomes covalently bound to endothelium and basement membrane, after induction of the classical antibody-induced pathway. As an established marker of antibody-mediated acute renal allograft rejection and its proclivity for endothelium, this component can be detected in peritubular capillaries in both chronic renal allograft rejection as well as hyperacute rejection, acute vascular rejection, acute cellular rejection, and borderline rejection. It has been shown to be a significant predictor of transplant kidney graft survival and is an aid in treating acute rejection.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Jianghua C, et al. Clin Transplant. 2005; 19:785-91
References 2:
Kayler LK, et al. Transplantation. 2008; 85:813-20
References 3:
Ranjan P, et al. Nephrol Dial Transplant .2008; 23:1735-41
References 4:
Seemayer CA, et al. Nephrol Dial Transplant. 2007; 22:568-76
References 5:
Bouron-Dal Soglio D, et al. Hum Pathol. 2008; 39:1103-10
SV40, Simian Virus 40 is a polyomavirus that is found in both monkeys and humans. Like other polyomaviruses, SV40 is a DNA virus that has the potential to cause tumors. SV40 is believed to suppress the transcriptional properties of tumor-suppressing p53 in humans through the SV40 large T-antigen and SV40 small T-antigen. It is generally assumed that large T-antigen is the major protein involved in neoplastic processes and the large T-antigen predominantly exerts its effect through deregulation of tumor suppressor p53, which is responsible for initiating regulated cell death (apoptosis), or cell cycle arrest when a cell is damaged. A mutated p53 gene may contribute to uncontrolled cellular proliferation, leading to a tumor.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gurney, E.G., et al. J Virl. 34:752-763 (1980)
References 2:
Huang, H., Reis,R. et al. Brain Pathol., 9:33-42 (1999)
References 3:
Arrington, A.S., et al. Molecular and Clinical Perspectives; 461-489 (2001)
Annexin A1, also known as lipocortin I, is a protein that is encoded by the ANXA1 gene in humans. Annexin A1 is a useful marker for identifying hairy cell leukemia cells. ANXA1 is strongly expressed on the cell membrane and occasionally in the cytoplasm of tumor cells in 97% of samples from patients with hairy cell leukemia, it is therefore a useful marker for identifying hairy cell leukemia cells. By contrast, B-cell lymphomas other than hairy cell leukemia, including typical splenic lymphoma with villous lymphocytes and patients with variant hairy cell leukemiaas defined by current morphologic, phenotypic, and clinical criteriaare ANXA1-negative. Additionally, aberrant expression of Annexin A1 has been reported in certain types of breast and gastric carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Falini B, et al. Lancet.; 363:1869-70 (2004)
References 2:
Sobral-Leite M, et al. BMC Med.; 13:156 (2015)
References 3:
Cheng TY, et al. Cancer.; 118:5757-67 (2012)
References 4:
Sato Y, et al. Exp Ther Med.; 2:239-43 (2011)
References 5:
Wang KL, et al.. Clin Cancer Res.; 12:4598-604 (2006)
Wilms tumor 1 protein (WT1) is a zinc finger transcription factor, normally expressed in tissues of mesodermal origin. The Wilms tumor gene encodes a protein that functions as a tumor suppressor gene. WT1 is detected in tumor cells of Wilms Tumor (also known as nephroblastoma) and mesothelioma. Additionally, WT1 expression has been found in ovarian serous carcinomas and some breast carcinomas.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
6F-H2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Charles AK; Moore IE; Berry PJ. Histopathology ; 30(4):312-4 (1997)
References 2:
Ordonez NG. Am J Surg Pathol 24(4):598-606, (2000)
References 3:
Foster MR, et al. Arch Pathol Lab Med; 125:1316-20 (2001)
References 4:
Nakatsuka S, et al. Mod Pathol; 19:804-14 (2006)
References 5:
May RJ, et al. Clin Cancer Res.; 13: 4547-55 (2007)
Vimentin is a 57kD, class-III, intermediate filament found in a variety of mesenchymal cells, including melanocytes, lymphocytes, endothelial cells, and fibroblasts. Non-reactivity of anti-vimentin is often considered more useful than its positive reactivity, since there are a few tumors that do not contain vimentin. The anti-vimentin immunohistochemical reagent is also useful as a tissue process control reagent.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
V9
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ishii Y, et al. Clin Exp Immunol. 1984; 58:183-92
References 2:
Battifora H. Am J Clin Pathol. 1991; 96:669-71
References 3:
Takeyoshi I, et al. Hepatogastroenterology. 2000; 47:1611-4
References 4:
Yaziji H, et al. Int J Gynecol Pathol. 2001; 20:64-78
Villin is an actin-binding glycoprotein that serves an important role in the maintenance of the microvilli brush border in gastrointestinal (GI) mucosal epithelium and its associated tumors. Recent immunohistochemical studies with villin have shown that villin is not only expressed in carcinomas of the gastrointestinal tract, but also in renal cell carcinomas, pancreatic carcinomas, endometrial carcinomas, as well as carcinomas of the ovary and lungs. In addition, positive villin expression may be seen in neuroendocrine/carcinoid tumors of the GI tract and lungs.
Antibody Isotype:
N/A
Monosan Range:
MONOSAN Ready To Use
Clone:
CWWB1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Robert W. Werling et al. Am J Surg. Path.2003; 27(3):303-310
References 2:
Tamboli P. et al Arch Pathol Lab Med 2002;126:1057-1063
References 3:
Zhang P. J. et al. Arch Pathol Lab Med. 1999;123:812-816
References 4:
Nishizuka S et al. Cancer Res. 2003 Sep 1;63(17):5243-50
Uroplakins (UPs) are a family of transmembrane proteins (UPs Ia, Ib, II and III) that are specific differentiation products of urothelial cells. In non-neoplastic mammalian urothelium, UPs are expressed in the luminal surface plasmalemma of superficial (umbrella) cells, forming complexes of 16nm crystalline particles. UPIII is specific for tumors of urothelial origin and, when used in combination with other markers, can aid in the diagnosis of primary and metastatic tumors.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
AU-1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Moll R, et al. Am J Pathol. 1995; 147:1383-97
References 2:
Olsburgh J, et al. J Pathol. 2003; 199:41-9
References 3:
Parker DC, et al. Am J Surg Pathol. 2003; 27:1-10
References 4:
Ohtsuka Y, et al. BJU Int. 2006; 97:1322-6
References 5:
Logani S, et al. Am J Surg Pathol. 2003 Nov;27(11):1434-41
Tyrosinase is an enzyme, amongst a family of enzymes, which is involved in the biosynthesis of melanin. It is a highly specific and sensitive marker for melanocytic differentiation, and has been found to be quite specific for melanotic lesions such as malignant melanoma.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
T311
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kaufmann O, et al. Mod Pathol 1998 Aug; 11(8):740-6
References 2:
Meije CB; et al. J Pathol 2000 Apr; 190(5):572-8
References 3:
Kanitakis J et al. Am J Dermatopathol. 2002 Dec;24(6):498-501
References 4:
Eudy GE et al. Hum Pathol. 2003 Aug;34(8):797-802
References 5:
Jaanson N et al. Melanoma Res. 2003 Oct;13(5):473-82
Anti-TTF-1 (Thyroid Transcription Factor 1) is useful in differentiating primary adenocarcinoma of the lung from metastatic carcinomas originating in the organs rather than thyroid, germ cell tumors, and malignant mesothelioma. It can also be used to differentiate small cell lung carcinoma from lymphoid infiltrates. TTF-1 labeling is also seen in thyroid and thyroid-derived tumors. TTF-1 immunostaining is useful in the differentiation between pulmonary and nonpulmonary origin of adenocarcinomas in malignant effusions. TTF-1 staining is very reliable in discerning whether a brain metastasis has arisen from a pulmonary or nonpulmonary site, particularly when dealing with adenocarcinomas and largecell carcinomas.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
8G7G3/1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Bejarano PA, et al. Mod Pathol. 1996; 9:445-52
References 2:
Di Loreto C, et al. Cancer Lett. 1998; 124:73-8
References 3:
Abutaily AS, et al. J Clin Pathol. 2002; 55:662-8
References 4:
Jang KY, et al. Anal Quant Cytol Histol. 2001; 23:400-4
Thyroid-stimulating hormone (also known as TSH or thyrotropin) is a peptide hormone synthesized and secreted by thyrotrops in the anterior pituitary gland which regulate the endocrine function of the thyroid gland. TSH is a glycoprotein and consists of two subunits which are non-covalently bound to one another. Anti-TSH reacts with TSH-producing cells (thyrotrophs), and is a useful marker in classification of pituitary tumors.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Batanero E, et al. Brain Behav Immun. 1992; 6:249-64
References 2:
Sanno N, et al. J Clin Endocrinol Metab. 1995; 80:2518-22
References 3:
La Rosa S, et al. Virchows Arch. 2000; 437:264-9
References 4:
Kuzuya N, et al. J Clin Endocrinol Metab. 1990; 71:1103-11
Tryptases compose a subfamily of proteinases with trypsin-like activity that are mostly stored in mast cell secretory granules and released into the extracellular environment upon mast cell activation. Several biological functions for tryptases have been proposed, including involvement in inflammatory and allergic responses.1 Mature mast cells have a complex distribution throughout the body. Antitryptase is a useful marker for mast cells.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
G3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Jordan JH et al. Hum Pathol. 2001 May;32(5):545-52
References 2:
Gordon LK et al. Clin Immunol. 2000 Jan;94(1):42-50
References 3:
Ghott A et al. Am J Surg Pathol. 2003 Jul;27(7):1013-9
References 4:
Aoki M et al. Int Arch Allergy Immunol. 2003 Mar;130(3):216-23
References 5:
Fiorucci L, et al. Cell Mol Life Sci. 2004 Jun;61(11):1278-95
Toxoplasma gondii is a spindle-to-oval-shaped protozoan which presents as an infection in humans of various sorts. The cyst (30 um) and trophozoite (7 um) stages can be identified in humans is such cases. This intracellular parasite is transmitted via raw/undercooked meat, contaminated soil, or by direct contact with an infected host. Infection in humans is usually associated with a variable degree of immunosuppression such as in pregnancy or immunosuppression due to various drugs.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Thyroglobulin (Tg) is the precursor of the iodinated thyroid hormones thyroxine (T4) and triiodothyronine (T3). Tg is a high molecular weight glycoprotein found in normal thyroid follicular cells. Thyroglobulin is useful for identifying thyroid carcinoma of papillary and follicular types and for identifying tumors of thyroid origin when working with adenocarcinoma of unknown primary.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
2H11+6E1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sellitti DF and Suzuki K. Thyroid. 2014; 24:625-38
References 2:
Bellet D, et al. J Clin Endocrinol Metab 1983; 56:530-3
References 3:
Bejarano PA, et al. Appl Immunohistochem Mol Morphol. 2000; 8:189-94
Anti-TdT antibody labels normal cortical thymocytes and primitive lymphocytes. Anti-TdT antibody detects an enzyme found in the nucleus of normal hematopoietic cells, normal cortical thymocytes and in the cytoplasm of megakaryocytes of the bone marrow. TdT expression is seen in over 90% of acute lymphoblastic lymphoma/ leukemia cases with the exception of pre-B-Cell ALL. TdT expression is not seen in normal mature T-or B-lymphocytes. Anti-TdT is positive for approximately one third of all cases of chronic myeloid leukemia, making it a good indicator of better response to chemotherapy.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Motea EA, et al. Biochimica et Biophysica Acta. 2010; 1804:1151-66
References 2:
Stauchen JA, et al. Int J Surg Pathol. 2003; 11:21-4
References 3:
Suzumiya J, et al. J Pathol. 1997; 182:86-91
References 4:
Arber DA, et al. Am J Clin Pathol. 1996; 106:462-8
Tumor associated glycoprotein (TAG)-72 is a high molecular weight glycoprotein that is present on the surface of many neoplastic cells, including adenocarcinomas of the breast, colon, and lung. TAG-72 is found in lung adenocarcinoma and is absent in mesothelioma, making the TAG-72 antibody useful in distinguishing adenocarcinoma from mesothelioma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
B72.3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Thor, A, et al. Cancer Res 1986;46:3118
References 2:
Schlom J, et al. Tumormarker Oncology;1987;2:3
References 3:
Osteen KG et al. In J Gynecol Pathol. 1992 Jul;11(3):216-20
References 4:
Ordonez NG. Am J Surg Pathol. 1998 Oct;22(10):1203-14
References 5:
Chhieng DC et al. Hum Pathol. 2003 Oct;34(10):1016-21
Anti-Synaptophysin reacts with neuroendocrine cells of human adrenal medulla, carotid body, skin, pituitary, thyroid, lung, pancreas and gastrointestinal mucosa. Positive staining is seen in neurons of the brain, spinal cord, retina, and Paneths cells in the gastrointestinal tract and gastric parietal cells. This antibody identifies normal neuroendocrine cells and neuroendocrine neoplasms. Diffuse, finely granular cytoplasmic staining is observed, which probably correlates with the distribution of the antigen within neurosecretory vesicles. The expression of synaptophysin is independent of the presence of NSE or other neuroendocrine markers. Anti-Synaptophysin is an independent broadrange marker of neural and neuroendocrine differentiation.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Navone F, et al. J Cell Biol. 1986; 103:2511-27
References 2:
Wiedenmann B, et al. Cell. 1985; 41:1017-28
References 3:
Kayser K, et al. Pathol Res Pract. 1988; 183:412-7
Spectrin is a cytoskeletal protein which is found in muscles, red blood cells and red cell precursors. Anti-Spectrin antibody is useful in the identification of blood dyscrasias and muscle disorders.
Antibody Isotype:
IgG2b-k
Monosan Range:
MONOSAN Ready To Use
Clone:
RBC2/3D5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sadahira, Y et al. J Clin Pathol. 1999 Dec; 52(12): 919-21
References 2:
Nehls, V et al. Am J Pathol. 1993 May; 142(5): 1565-73
References 3:
Muller M et al. J Vet Med A Physiol Pathol Clin Med. 2001 Feb;48(1):51-7
References 4:
Terada N et al. J Anat. 1997 Apr;190(Pt 3):397-404
Smooth Muscle Myosin, heavy chain (SMMS-1) is a cytoplasmic structural protein that is a major component of the contractile apparatus of the smooth muscle cells. SMMS-1 is also a myoepitheliumassociated protein. Anti-SMMS-1 is a mouse monoclonal antibody to smooth muscle myosin, heavy chain that reacts with human visceral and vascular smooth muscle cells. The antibody also reacts with human myoepithelial cells. It is very helpful in distinguishing between benign sclerosing breast lesions and infiltrating carcinomas in difficult cases since it strongly stains the myoepithelial layer in the benign lesions while it is negative in the infiltrating carcinomas.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
SMMS-1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Werling RW, et al. Am J Surg Pathol. 2003; 27:82-90
References 2:
Agoff SN, et al. Appl Immunohistochem Mol Morphol. 2001; 9:164-9
References 3:
Popnikolov NK, et al. Am J Clin Pathol. 2003; 120:161-7
References 4:
Lazard D, et al. Proc Natl Acad Sci USA. 1993; 90:999-1003
S-100 protein has been found in normal melanocytes, Langerhans cells, histiocytes, chondrocytes, lipocytes, skeletal and cardiac muscle, Schwann cells, epithelial and myoepithelial cells of the breast, salivary and sweat glands, as well as in glial cells.1,2,6 Neoplasms derived from these cells also express S-100 protein, albeit non-uniformly.1-4 A large number of well differentiated tumors of the salivary gland, adipose and cartilaginous tissue, 3 and Schwann cell-derived tumors express S-100 protein. Almost all malignant melanomas and cases of histiocytosis X are positive for S-100 protein.4,5 Despite the fact that S-100 protein is an ubiquitous substance, its demonstration is of great value in the identification of several neoplasms, particularly melanomas.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
4C4.9
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Nakajima T, et al. Ad J Surg Path. 1982; 6:715-727
References 2:
Kuhn HJ, et al. Am J Clin Path. 1983; 79:341-347
References 3:
Yaziji H, et al. Int J Surg Pathol. 2003; 11:11-5
References 4:
Patel P, et al. J Am Acad Dermatol. 2002; 46:264-70
References 5:
Morrison CD, et al. Semin Diagn Pathol. 2000; 17:204-15
Anti-renal cell carcinoma (RCC) recognizes a 200 kD glycoprotein localized in the brush border of the proximal renal tubule. This antibody immunoreacts with approximately most primary renal cell carcinomas and can aid in the diagnosis when renal cell carcinoma enters the differential diagnosis. Other tumors that may react with this antibody are parathyroid adenoma, an occasional breast carcinoma. Nephroblastoma, oncocytoma, mesoblastic nephroma, transitional cell carcinoma, and angiomyolipoma are not labeled with this antibody.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
PN-15
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Dabbs, D. 4th Edition. Elsevier Saunders. 2014; p234
References 2:
Bakshi N, et al. Appl Immunohistochem Mol Morphol. 2007; 15:310-5
References 3:
McGregor DK, et al. Am J Surg Pathol. 2001; 25:1485-92
References 4:
Avery, AK et al. Am J Surg Pathol 24(2): 203-210, 2000
The antibody reacts with prostatic acid phosphatase in the glandular epithelium of normal and hyperplastic prostate, and adenocarcinoma of the prostate. Anti-PSAP is useful in identifying prostatic origin of tumors in the metastatic setting. PSAP complements other immunohistochemical markers in the correct clinical context.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
PASE/4LJ
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ansari, MA, et al. Am J Clin Path 1981;76:94-98
References 2:
Kimura, N, et al. Virchows Arch A 1986;4:247-251
References 3:
Kidwai N et al. Breast Cancer Res. 2004;6(1):R18-23
References 4:
Genega EM et al. Mod Pathol. 2000 Nov;13(11):1186-91
References 5:
Gatalica Z et al. Appl Immunohistochem Mol Morphol. 2000 Jun;8(2):158-61
Prostate-Specific Antigen (PSA) is a 33 kDa protein primarily produced by the prostatic epithelium and the epithelial lining of the periurethral glands. PSA is strongly expressed in both normal and neoplastic prostatic tissue. Although PSA can be considered prostate-specific, PSA and/or PSA gene expression has been detected at low levels in some extra-prostatic tissues such as normal breast tissue, breast tumors, endometrium, adrenal neoplasms and renal cell carcinomas. Anti-PSA is most useful in determining the prostatic origin of carcinomas in non-prostate tissues (metastatic disease) using IHC techniques. This product is best used in conjunction with a panel of antibodies as, up to 27% of prostate carcinoma cases (predominantly poorly differentiated carcinomas) can be negative for this marker.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
ER-PR8
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Polascik TJ, et al. J Urol. 1999; 162:293
References 2:
Stenman UH, et al. Semin Cancer Biol. 1999; 9:83-93
References 3:
Alanen KA, et al. Path Res Pract. 1996; 192:233-237
Prolactin (PRL) is a single-chain polypeptide of 226 amino acids and plays a role in multiple processes including cell growth, reproduction, and immune function. Anti-Prolactin reacts with prolactin-producing cells and is a useful marker in classification of pituitary tumors and the study of pituitary disease.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Asa SL, et al. Arch Pathol Lab Med. 1982; 106:360-3
References 2:
Duello TM, et al. Am J Anat. 1980; 158:463-9
References 3:
Minniti G, et al. Surg Neurol. 2002; 57:99-103
References 4:
Popadic A, et al. Surg Neurol. 1999; 51:47-54
References 5:
Nevalainen MT, et al. J Clin Invest. 1997; 99:618-27
The antibody reacts with progesterone receptor forms alpha and beta. This antibody stains nuclei in breast, ovarian and endometrial epithelia, as well as myometrial nuclei. Since the early 1990s the immunohistochemical (IHC) assay determination of progesterone receptor status has replaced the dextran-coated charcoal method as a prognostic indicator in breast carcinoma. IHC has shown to be superior in prognostic significance when using any one of several available methods of quantitation using this technique.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
Y85
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Dabbs D. Diagnostic Immunohistochemistry, 2nd edition. p 728-32
References 2:
Dunnwald LK, et al. Breast Cancer Res. 2007;9(1):R6
References 3:
Leong A S-Y, et al. Manual of diagnostic immunohistochemistry, 2nd edition. p 375-76
Pneumocystis carinii is a fungal organism which is detected in human tissues (typically in lung in immunocompromised patients) in the trophozoite stage. Anti-Pneumocystis carinii antibody reacts with an epitope on the organism which is resistant to formalin, picric acid, paraffin, as well as alchohol and xylene. No cross-reactivity has been demonstrated with other fungi or parasitic organisms.
Antibody Isotype:
IgM
Monosan Range:
MONOSAN Ready To Use
Clone:
3F6
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Anti-PLAP antibody immunoreacts with germ cell tumors and can discriminate between these and other neoplasms. Somatic neoplasms e.g. breast, gastrointestinal, prostatic and urinary cancers may also immunoreact with antibodies to PLAP. Anti-PLAP positivity in conjunction with keratin negativity favors seminoma over carcinoma. Germ cell tumors are usually keratin positive, but they regularly fail to stain with anti-EMA, whereas most carcinomas stain with anti-EMA. Anti-PLAP has been useful in the diagnosis of gestational trophoblastic disease. This antibody has shown cross-reaction with human intestinal alkaline phosphatase.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
NB-10
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Paiva, J, et al. Am J Pathol 1984;111:156-165
References 2:
Burke, AP, et al. Hum Path 1988;19:663-670
References 3:
Wick, MR, et al. Hum Path 1987;18:946-954
References 4:
Saad RS et al. Appl Immunohistochem Mol Morphol. 2003 Jun;11(2):107-12
References 5:
Goldsmith JD et al. Am J Surg Pathol. 2002 Dec;26(12):1627-33
Protein gene product 9.5 (PGP 9.5), also known as ubiquitin carboxyl-terminal hydrolase-1 (UCH-L1), is a 27-kDa protein originally isolated from whole brain extracts (1). Although PGP9.5 expression in normal tissues was originally felt to be strictly confined to neurons and neuroendocrine cells (2), it has been subsequently documented in distal renal tubular epithelium, spermatogonia, Leydig cells, oocytes, melanocytes, prostatic secretory epithelium, ejaculatory duct cells, epididymis, mammary epithelial cells, Merkel cells, and dermal fibroblasts. LK Campbell et al demonstrated immunostaining of a plethora of different mesenchymal neoplasms with this antibody.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Campbell LK, et al. Mod Pathol. 2003; 16:963-9
References 2:
Bassotti G, et al. J Clin Pathol. 2005; 58:973-7
References 3:
Mahalingam M, et al. J Cutan Pathol. 2001; 28:282-6.
References 4:
Mahalingam M, et al. J Cutan Pathol. 2006; 33:51-6.
Perforin is a pore-forming protein that leads to osmotic lysis of the target cells and subsequently enables granzymes to enter the target cells and activate apoptosis, the cell death program. The expression of perforin is reportedly upregulated in activated CD8+ T-cells, natural killer cells and some CD4+ T-cells.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-23
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Chu PG, et al. Ann Diagn Pathol. 1999; 3:104-33
References 2:
Bittmann I, et al. Arch. 2004; 445:375-81
References 3:
dAmore ES, et al. Pediatr Dev Pathol. 2007; 10:181-91
Programmed death-1 (PD-1), also known as CD279, is a type-I transmembrane protein expressed on T-cells, B-cells, and monocytes during activation.1-2 PD-1 and its ligands (PD-L1 and PD-L2) function as an immune checkpoint pathway by regulating T-cell activation and autoimmunity.2 PD-1 labels follicular helper T-cells and is therefore a useful marker for angioimmunoblastic T-cell lymphoma.3-4
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
NAT105
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Bolstad AI, et al. Arthritis Rheum. 2003 Jan;48(1):174-85
References 2:
Dorfman DM, et al. Am J Surg Pathol. 2006 Jul;30(7):802-10
References 3:
Hamanishi J, et al. Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3360-5
References 4:
Konishi J, et al. Clin Cancer Res. 2004 Aug 1;10(15):5094-100
References 5:
Mataki N, et al. Am J Gastroenterol. 2007 Feb;102(2):302-12
PAX-5 encodes for B-cell-specific activator protein (BSAP), a marker for B-cells, including B-lymphoblastic neoplasms and maturation stage. It is found in most cases of mature and precursor B-cell non-Hodgkin lymphomas/leukemias. In approximately 97% of cases of classic Hodgkin lymphoma, Reed-Sternberg cells express PAX-5.4 PAX-5 is not detected in multiple myeloma and solitary plasmacytoma, making it useful for such differentiation.1,3 Diffuse large B-cell lymphomas do express PAX-5, save for those with terminal B-cell differentiation. T-cell neoplasms do not stain with anti-PAX-5. There is a strong association with CD20 expression.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
24
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Torlakovic E, et al. Am J Surg Pathol. 2002; 26:1343-50
The antibody targets the capsid proteins VP1 and VP2 on Human Parvovirus. Parvovirus B19 infection has been implicated as a cause in spontaneous abortion in humans and thus application of this antibody to placental tissues in such cases is appropriate. Parvovirus B19 is also associated with erythema infectiosum (fifth disease) in children and acute arthritis in adults, as well as chronic hemolytic anemia, with some patients experiencing aplastic crisis.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
R92F6
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Loughrey AC et al. J Med Vir 39:97-100 (1993)
References 2:
Moore L et al. Med J Australia 159:344-345 (1993)
References 3:
Morey AL et al. J Path 166:105-108 (1992)
References 4:
ONeill HJ et al. 123: 125-134 (1992)
References 5:
Silverberg SG et al. Principles and Practice of Surgical Pathology and Cytopathology, 3rd edition. 1997; p. 219-220
The antibody has been used as an aid in discriminating complete hydatidiform mole (CHM) (no nuclear labeling of cytotrophoblasts or stromal cells) from partial hydatidiform mole (PHM) (nuclear staining of both cytotrophoblasts and stromal cells) and hydropic abortion. In normal placenta, cytotrophoblast, syncytio trophoblast, and stromal cells are labeled with this antibody. Intervillous trophoblastic islands demonstrate nuclear labeling in all entities and serve as an internal control.
Antibody Isotype:
IgG2b-k
Monosan Range:
MONOSAN Ready To Use
Clone:
KP10
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kihara M, et al. J Reprod Med. 2005; 50:307-12
References 2:
Romaguera RL, et al. Fetal Pediatr Pathol. 2004; 23:181-90
The antibody recognizes a 53 kDa phosphoprotein, identified as p53 suppressor gene product. It reacts with the mutant as well as wild type p53.1 Positive nuclear staining with this antibody has been shown to be a factor in breast carcinoma, lung carcinoma, colorectal carcinoma, urothelial carcinoma, and ependymoma.2-8 Anti-p53 positivity has also been used to differentiate uterine serous carcinoma from endometrioid carcinoma, as well as a marker for intratubular germ cell neoplasia.
Antibody Isotype:
IgG2b-k
Monosan Range:
MONOSAN Ready To Use
Clone:
DO7
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mauri FA et al. Int J Oncol 1999 Dec;15(6):1137-47
References 2:
Caffo O et al. Clin Cancer Res 1996 Sep;2(9):1591-9
References 3:
Bebenek M et al. Anticancer Res 1998 Jan-Feb;18(1B):619-23
References 4:
Midulla C et al. Anticancer Res 1999 Sep-Oct;19(5B):4033-7
References 5:
Moore BE et al. App.IHC and Mol. Morphol. 2001;9(3): 203 206
Alpha-Methylacyl-CoA Racemase (also known as AMACR or P504s) is an essential enzyme in the ?oxidation of branched-chain fatty acids. AMACR over-expression has been demonstrated in several cancers including colorectal, prostate, ovarian, breast, bladder, lung, and renal cell carcinomas, lymphoma, and melanoma. Staining with the antibody to this enzyme has been useful in identifying prostate carcinoma and prostatic intraepithelial neoplasia, as well as atypical adenomatous hyperplasia in formalin-fixed paraffinized tissue in morphologically difficult cases.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
13H4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Browne TJ et al. Hum Pathol. 2004 Dec;35(12):1462-8
References 2:
Wu CL et al. Hum Pathol. 2004 Aug;35(8):1008-13
References 3:
Evans AJ. J Clin Pthol. 2003 Dec;56(12):892-7
References 4:
Beach R, Gown AM, et al. Am J Surg Pathol. 2002 Dec;26(12):1588-96
References 5:
Jiang Z, Wu CL, et al. Am J Surg Pathol. 2002 Sep;26(9):1169-74
p27, also known as cyclin-dependent kinase inhibitor 1B (CDNK1B), is a kinase inhibitor that controls cell cycle progression.1-4 p27 is involved in G1 phase arrest and obstructs cell entry into the S phase by binding to and inhibiting cyclin E-CDK2, effectively slowing or stopping the cell division cycle.1-4 p27 is broadly expressed in normal tissue but can be dysfunctional in neoplastic tissue and, therefore, not expressed.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
SX53G8
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Oct-4 is a transcription factor that functions in the regulation and maintenance of pluripotency in embryonic stem and primordial germ cells. Oct-4 immunoreactivity has been demonstrated in gonadal and extra-gonadal seminomas, dysgerminomas and embryonal carcinomas. In addition, the immunohistochemical detection of Oct-4 assists in the evaluation of intratubular germ cell neoplasia (IGCN).
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-10
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Cheng L, et al. J Pathol. 2007; 211:1-9
References 2:
Weissferdt A, et al. Hum Pathol. 2015; 46:376-83
References 3:
Browne P, et al. Am J Clin Pathol. 2003 Nov;120(5):767-77
References 4:
García-Cosío M, et al. Mod Pathol. 2004 Dec;17(12):1531-8
References 5:
Gibson SE, et al. Am J Clin Pathol. 2006 Dec;126(6):916-24
Nerve growth factor receptor (NGFR), also known as p75NTR, is a 75-kDa glycoprotein member of the tumor necrosis factor (TNF) receptor family essential for embryonic development of the peripheral nervous system. In normal tissue, NGFR is expressed in neural crest derived cells, lymphoid follicular dendritic cells seen in lymph nodes and tonsils, and myoepithelial cells of the breast, prostate, and salivary gland as well as basal epithelium of the respiratory system. NGFR has been shown to be a reliable adjunct marker for melanoma, specifically desmoplastic and spindle cell variants Anti-NGFR labels myoepithelial cells of the breast and may aid in the differentiation between benign conditions, pre-invasive neoplastic lesions and invasive malignancies of the breast.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-21
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Thompson SJ. Am J Clin Pathol. 1989; 92:415-23
References 2:
Reis-Filho JS, et al. Mod Pathol. 2006; 19:307-19
References 3:
Lazova R, et al. J Am Acad Dermatol. 2010; 63:852-8
Neurofilaments (NF) are intermediate filaments that provide structural support to the neuronal cytoskeleton.1-3 They are heteropolymers composed of three NF subunits: NF-H, NF-M, and NF-L.2 Anti-NF labels neurons of the central and peripheral nervous system as well as neoplastic cells of neuronal origin.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
2F11
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Diepholder HM, et al. Cancer. 1991; 68:2192-201
References 2:
Lee M, et al. J. Cell Biol. 1993; 122:1337-50
References 3:
Trojanowski JQ, et al. Hum Pathol. 1984; 15:248-57
Immunostaining with anti-myoglobin provides a specific, sensitive, and practical procedure for the identification of tumors of muscle origin. Since myoglobin is found exclusively in skeletal and cardiac muscle and is not present in any other cells of the human body, it may be used to distinguish rhabdomyosarcoma from other soft tissue tumors.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mukai K, et al. Am J Surg Pathol. 1979; 3:373-6
References 2:
Corson JM, et al.Am J Pathol. 1981; 103:384-9
References 3:
Brooks JJ. Cancer. 1982; 50:1757-63
References 4:
Furlong MA, et al. Ann Diagn Pathol. 2001; 5:199-206
Myogenin also identified as myogenic factor 4 is a muscle specific transcription factor associated with muscle differentiation and cell cycle.1 Anti-myogenin reactivity is seen in the nuclei of myoblasts in developing muscle tissue. Anti-myogenin is a useful immunohistochemical reagent for identification of rhabdomyosarcoma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
F5D
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Miller JB. J Cell Biol.; 111:1149-59 (1990)
References 2:
Wang NP, et al. Am J Pathol.; 147:1799-810 (1995)
References 3:
Cui S, et al. Pathol Int.; 49:62-8 (1999)
References 4:
Kaspar et al. Sci Rep.; 5:15090 (2015)
References 5:
Rudzinski et al. Am J Surg Pathol.; 38:654-9 (2014)
Anti-Myeloperoxidase detects granulocytes and monocytes in blood and precursors of granulocytes in the bone marrow. This antibody can detect myeloid cell populations of the bone marrow as well as in other sites.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Pinkus GS, et al. Mod Pathol. 1991; 4:733-41
References 2:
Markoc F, et al. Tumori. 2010; 96:149-53
References 3:
Alexiev BA, et al. Diagn Pathol. 2007; 31;2:42
References 4:
Saravanan L, et al. Int J Lab Hematol. 2010; 32:132-6
References 5:
Manaloor EJ, et al. Am J Clin Pathol. 2000; 113:814-22
Mucins are high molecular weight glycoproteins which constitute the major component of the mucus layer that protects the gastric epithelium from chemical and mechanical aggressions. In humans, at least 14 mucin genes have been identified that code for the mucin proteins. MUC6 is a secretory mucin that is part of a family of at least 14 high molecular weight glycoproteins made by many epithelial tissues. MUC6 is preferentially expressed in non-neoplastic gastric tissue, specifically in the pyloric glands. During neoplastic transformation, mucin expression may be altered within these tissues leading to particular patterns of expression.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-20
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ruco LP, et al., Am J Clin Pathol. 92:273-9 (1989)
References 2:
Facchetti F, et al., Histopathology. 19:141-5 (1991)
References 3:
Do SI. et al. J of Breast Cancer. 16:152-8 (2013)
References 4:
Vergier B et al. Blood., 9597: 2212-8 (2000)
References 5:
Mino-Kenudson M, et al. Virchows Arch. 469:255-65 (2016)
Mucins are high molecular weight glycoproteins which constitute the major component of the mucus layer that protects the gastric epithelium from chemical and mechanical injury. In humans, at least 14 mucin genes have been identified that code for the mucin proteins. Reportedly, mucin expression is associated with tumor type of gastric carcinomas, with MUC2 being associated with mucinous carcinomas. Anti- MUC2 reactivity is seen in goblet cells of the small intestine and colon, and it is useful in immunohistochemistry for identifying colonic, gastric and esophageal carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-18
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Chaves P, et al. Dis Esophagus. 2005; 18:383-7
References 2:
Leteurtre E, et al. World J Gastroenterol. 2006; 12:3324-31.
References 3:
Mino-Kenudson M, et al. Arch Pathol Lab Med. 2007; 131:86-90
References 4:
Mizoshita T, et al. Histol Histopathol. 2007; 22:251-60
References 5:
OConnell FP, et al. Arch Pathol Lab Med. 2005; 129:338-47
Mucins are high molecular weight glycoproteins which constitute the major component of the mucus layer that protects the gastric epithelium from chemical and mechanical injury. In humans, at least 14 mucin genes have been identified that code for the mucin proteins. Mucin genes are expressed in a regulated cell- and tissue-specific manner. The stomach provides a good example of such differential expression of mucin genes. MUC1 is detected in mucous cells of the surface epithelium and neck region of the gastric antrum, as well as in pyloric glands and oxyntic glands of the body region. The heterogeneous pattern of mucin expression, including the expression of the intestinal mucin MUC2, may provide new insights into the differentiation pathways of gastric carcinoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-17
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Chaves P, et al. Dis Esophagus. 2005; 18:383-7
References 2:
Leteurtre E, et al. World J Gastroenterol. 2006; 12:3324-31.
References 3:
Mino-Kenudson M, et al. Arch Pathol Lab Med. 2007; 131:86-90
References 4:
Mizoshita T, et al. Histol Histopathol. 2007; 22:251-60
References 5:
OConnell FP, et al. Arch Pathol Lab Med. 2005; 129:338-47
MSH6 is a mismatch repair gene which is deficient in a high proportion of patients with microsatellite instability (MSI-H). This finding is associated with the autosomal dominant condition known as Hereditary Non-Polyposis Colon Cancer (HNPCC). The anti-MSH6 antibody is useful in screening patients and families for this condition. Colon cancers that are microsatellite unstable have a better prognosis than their microsatellite stable counterparts.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
44
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Lagerstedt Robinson K, et al. J Natl Cancer Inst. 2007 Feb 21;99(4): 291-9
References 2:
Niessen RC et al. Gut 2006 Dec;55(12):1781-8
References 3:
Lawes DA et al. Br J Cancer. 2005 Aug 22; 93(4):472-7
References 4:
Stormorken AT et al. J Clin Oncol. 2005 Jul 20;23(21):4705-12
References 5:
Rigau V et al. Arch Pathol Lab Med. 2003;127:694-700
MSH2 is a mismatch repair gene which is deficient in a high proportion of patients with microsatellite instability (MSI-H). This finding is associated with the autosomal dominant condition known as Hereditary Non-Polyposis Colon Cancer (HNPCC). The anti-MSH2 antibody is useful in screening patients and families for this condition. Colon cancers that are microsatellite unstable have a better prognosis than their microsatellite stable counterparts.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
G219-1129
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Christensen M et al. Cancer 2002;95: 2422-30
References 2:
Wright CL et al. Am J Surg Pathol. 2003;27: 1393-1406
References 3:
Renkonen E et al. J Clin Oncol 2003; 21: 3629-3637
References 4:
Hoedema R. et al. The American Surgeon 2003, May 69(5): 387-92
References 5:
Brueckl WM et al. Anticancer Research 2003; 23: 1773-1778
MLH1 is a mismatch repair gene that is deficient in a high proportion of patients with microsatellite instability (MSI-H). This finding is associated with the autosomal dominant condition known as Hereditary Non-Polyposis Colon Cancer (HNPCC). The anti-MLH1 antibody is useful in screening patients and families for this condition. Colon cancers that are microsatellite unstable have a better prognosis than their microsatellite stable counterparts
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
G168-728
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Christensen M et al. Cancer 2002;95: 2422-30
References 2:
Wright CL et al. Am J Surg Pathol. 2003;27: 1393-1406
References 3:
Renkonen E et al. J Clin Oncol 2003; 21: 3629-3637
References 4:
Hoedema R. et al. The American Surgeon 2003, May 69(5): 387-92
References 5:
Brueckl WM et al. Anticancer Research 2003; 23: 1773-1778
Hector Battifora mesothelial-1 (HBME-1) is a membrane antigen that exists in the microvilli of mesothelial cells and other epithelial cells. Anti-HBME-1 labels thyroid papillary carcinoma and follicular carcinoma but not normal thyroid making it a valuable marker for distinguishing thyroid malignacies from benign thyroid lesions. It has also been demonstrated to label mesothelial cells, both be
Antibody Isotype:
IgM
Monosan Range:
MONOSAN Ready To Use
Clone:
HBME-1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Coli A, Bigotti G, et al. J Exp Clin Cancer Res. 2007 Jun;26(2):221-7
Anti-HMB45 is a useful melanoma immunohistochemical marker that reacts with antigens present on immature melanosomes. Anti-HMB45 is useful for identifying amelanotic melanoma from other neoplastic lesions with similar morphology.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
HMB-45
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gown AM, et al. Am J Pathol. 1986; 123:195-203
References 2:
Wick MR, et al. Arch Pathol Lab Med. 1988; 112:616-20
References 3:
Abrahamsen HN, et al. Cancer. 2004; 100:1683-91
References 4:
Vaggelli L, et al. Tumori. 2000; 86:346-8
References 5:
Baisden BL, et al. Am J Surg Pathol. 2000; 24:1140-6
MART-1 (also known as Melan A) is a melanocyte differentiation antigen. It is present in melanocytes of normal skin, retina, nevi and in the majority of melanomas. Tyrosinase is an enzyme integral in the process of melanin synthesis, and found in most malignant melanomas. Therefore, this cocktail is useful for the identification of melanomas and melanocytic lesions
Antibody Isotype:
IgG2b/IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
M2-7C10 & T311
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Mammaglobin is breast-associated glycoprotein distantly related to secretoglobin family that includes human uteroglobin and lipophilin. Anti-mammaglobin labels cytoplasm of normal breast epithelial cells as well as primary and metastatic breast carcinomas. Absence of mammaglobin expression is typically seen in prostate, kidney, colon, rectum, small intestine, stomach, pancreas, lung and thyroid tissue.2,5 Mammaglobin may be used as part of an immunohistochemical panel for determination of metastatic breast carcinoma and tumor of unknown primary origin.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
31A5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Fleming TP et al. Ann NY Acad Sci 2000;923:78-89
References 2:
Bhargava R, et al. Am J Clin Pathol. 2007; 127:103-13
References 3:
Wang Z, et al. Int J Clin Exp Pathol. 2009; 2:384-9
References 4:
Han JH, et al. Arch Pathol Lab Med. 2003; 127:1330-4
Anti-Lysozyme stains myeloid cells, histiocytes, granulocytes, macrophages, and monocytes in human tonsil, colon and skin. It is an important marker that may demonstrate the myeloid or monocytic nature of acute leukemia. The restrictive nature of anti-lysozyme antibody staining suggests that lysozyme may be synthesized predominantly in reactive histiocytes rather than in resting, unstimulated phagocytes. Anti-lysozyme may aid in the identification of histiocytic neoplasias, large lymphocytes and classifying lymphoproliferative disorders.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Rehg J, et al. Toxicol Pathol. 2012; 40: 345-74
References 2:
Seifert RP, et al. Annals Diag Pathol. 2014; 18:253-60.
Luteinizing hormone (LH) is a heterodimeric glycoprotein produced by gonadotropic cells of the pituitary gland. Anti-LH is a useful marker to aid in the classification of pituitary tumors and the study of pituitary disease.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sano T, et al. Virchows Arch A Pathol Anat Histopathol. 1990; 417:361-7
References 2:
Felix I, et al. Hum Pathol. 1991; 22:719-21
References 3:
Saccomanno K, et al. J Clin Endocrinol Metab. 1994; 78:1103-7
The antibody detects surface immunoglobulin on normal and neoplastic B-cells. Anti-lambda staining is seen in B-cell follicles of human lymphoid tissue. When dealing with B-cell neoplasms, the determination of light chain ratios remains helpful. Most B-cell lymphomas express either kappa or lambda light chains, whereas reactive proliferations display a mixture of kappa and lambda positive cells. If only a single light chain type is detected, a lymphoproliferative disorder is very likely.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
Lamb14
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Hertel, BF, et al. Lab Invest 1977;36:12
References 2:
Taylor, CL Arch Pathol Lab Med 1978;12:113-121
References 3:
Abbondanzo SL et al. Ann Diagn Pathol. 1999 Oct;3(6):394
References 4:
Kurtin PJ et al. Am J Clin Pathol. 1999 Sep;112(3):319-29
References 5:
Ashton-Key M et al. Histopathology. 1996 Dec;29(6):525-31
The antibody detects surface immunoglobulin on normal and neoplastic B-cells. In paraffin-embedded tissue, anti-kappa exhibits strong staining of kappa-positive plasma cells and cells that have absorbed exogenous immunoglobulins. When dealing with B-cell neoplasms, the determination of light chain ratios remains the centerpiece. Most B-cell lymphomas express either kappa or lambda light chains, whereas reactive proliferations display a mixture of kappa and lambda positive cells. If only a single light chain type is detected, a lymphoproliferative disorder exists. Monoclonality is determined by a kappa-lambda ratio of greater than or equal to 3:1 or a lambda-kappa ratio greater than 2:1.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
L1C1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Hertel, BF, et al. Lab Invest 1977;36:12
References 2:
Mendes S, Dreno B. Acta Derm Venereol. 2003;83(3):167-70
References 3:
Lee LA et al. Am J Otolaryngol. 2002 Sep-Oct;23(5):316-20
References 4:
Taylor, CL Arch Pathol Lab Med 1978;12:113-121
References 5:
Schmid U et al. Am J Surg Pathol. 1995 Jan;19(1):12-20
The INI-1 gene, which encodes a functionally uncharacterized protein component of the hSWI/SNF chromatin remodeling complex, is often mutated or deleted in malignant rhabdoid tumor (MRT). Two isoforms of INI-1 that differ by the variable inclusion of amino acids are potentially produced by differential RNA splicing. The morphology of MRTs can present challenges in differential diagnosis. The overall survival of MRTs relative to its potential mimics [medulloblastoma, supratentorial primitive neuroectodermal tumors (sPNETs)] is quite low, and thus differentiation from these other tumors is desirable. Lack of nuclear labeling by anti-INI-1 is characteristic of MRT. The majority of medulloblastomas and sPNETs are labeled by anti-INI-1. MRTs also originate from the kidney and soft tissues.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-27
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Bourdeaut F, et al. J Pathol. 2007; 211:323-30
References 2:
Fowler DJ, et al. Fetal Pediatr Pathol. 2006; 25:159-68
References 3:
Haberler C, et al. Am J SurgPathol. 2006; 30:1462-8
References 4:
Janson K, et al. Pediatr Blood Cancer. 2006 Sep:47(3):279-84
Inhibin is a peptide hormone that inhibits FSH secretion from the pituitary. Inhibin is a dimer that consists of an alpha and beta subunit. In normal tissue, anti-inhibin alpha labels granulosa cells of the ovary, sertoli and leydig cells of the testis, and the zona reticularis of the adrenal cortex. Anti-inhibin alpha has demonstrated utility in the identification of sex cord stromal tumors and adrenal cortical tumors.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
R1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Stewart CJ, et al. Histopathology. 1997; 31:67-74
References 2:
McCluggage WG, et al. J Clin Pathol. 1998; 51: 114-6
References 3:
Kommoss F, et al. Mod Pathol. 1998; 11:656-64
References 4:
Guerrieri C, et al. Int J Gynecol Pathol. 1998; 17:266-71
Anti-IgM reacts with immunoglobulin mu (IgM) chains. IgM is one of the predominant surface immunoglobulins on B-lymphocytes. This antibody is useful when differentiating and sub-classifying hematolymphoid neoplasms.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Arnold A, et al. New Eng J Med. 1983; 309:1593-1599
References 2:
Leong AS, et al. Geenwich Medical Media Ltd. 1999; 217-219
References 3:
Taylor CR. Arch Path Lab Med. 1978; 102:113-121
References 4:
Kojima M, et al. APMIS. 2002; 110:875-80
References 5:
Pambuccian SE, et al. Am J Surg Pathol. 1997; 21:179-86
Immunoglobulin A (IgA) plays a critical role in mucosal immunity. It is present in the mucosal secretions such as tears, saliva, colostrum, intestinal juice, vaginal fluid, and secretions from the prostate and respiratory epithelium, and represents a key first line of defense against invasion by inhaled and ingested pathogens at the vulnerable mucosal surfaces. It is also found in small amounts in blood. Because it is resistant to degradation by enzymes, secretory IgA can survive in harsh environments such as the digestive and respiratory tracts, to provide protection against microbes that multiply in body secretions. It is useful when identifying multiple myeloma.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ansari NA, et al. Asian Pac J Cancer Prev. 2007; 8:593-6
Human placental lactogen (hPL), also previously known as human chorionic somatomammotropin, is a 22-kD protein with partial homology to growth hormone. hPL is first detectable in the maternal serum in the fifth week of gestation and is involved in maintaining nutritient supply to the fetus. Anti-hPL reactivity is seen in syncytiotrophoblastic cells of placenta and choriocarcinoma
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Shih IM, et al. Am J Surg Pathol. 2004; 28:1177-83
References 2:
Ulbright TM, et al. Am J Surg Pathol. 1997; 21:282-8
Human herpesvirus type 8 (HHV-8) is the likely etiological agent of Kaposis sarcoma (KS). HHV-8 DNA sequences have been found in Kaposis sarcoma lesions, primary effusion lymphoma, and multicentric Castlemans disease via polymerase chain reaction and in situ hybridization. Latent nuclear antigen (LNA1, LNA, LANA-1), also known as ORF73, is a 222- or 234 kD protein that is consistently expressed in HHV8 infected cells. Anti-HHV-8 labels the latent nuclear antigen protein via immunohistochemistry
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
13B10
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Corbellino M et al AIDS Res Hum Retroviruses.;12(8):651-7 (1996)
References 2:
Schwartz EJ et al. Am J Surg Pathol.;27(12):1546-50 (2003)
References 3:
Boulanger E et al. Am J Hematol.;76(1):88-91 (2004)
References 4:
Courville P et al. Ann Pathol.;22(4):267-76 (2002)
Anti-hepatocyte specific antigen, also known as anti-Hep-Par1, recognizes both benign and malignant liver-derived tissues including such tumors as hepatocellular carcinoma. It recognizes both normal adult and fetal liver tissue. The typical pattern is a granular cytoplasmic staining. This antibody is useful in differentiating hepatocellular carcinomas with adenoid features from adenocarcinomas, either primary in the liver or metastatic lesions to the liver
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
OCH1E5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Minervini MI, et al. Mod Pathol. 1997; 10:686-92
References 2:
Chu PG, et al. Am J Surg Pathol. 2002; 26:978-88
References 3:
Wieczorek T, et al. Am J Clin Pathol. 2002; 118:911-21
References 4:
Fasano M, et al. Mod Pathol. 1998; 11:934-8
References 5:
Maitra A, et al. Am J Clin Pathol. 2001; 115:689-94
Helicobacter pylori is strongly associated with inflammation of the stomach and is also implicated in the development of gastric malignancy, peptic ulcers, and gastric lymphomas in humans. Helicobacter pylori can exist in a number of locations: in the mucus, attached to epithelial cells, or inside of vacuoles in epithelial cells, where it produces adhesions that bind to membrane-associated lipids and carbohydrates in or on epithelial cells. The most reliable method for detecting H. pylori infection is a biopsy during endoscopy histologic examination and detection by immunohistochemistry. Immunohistochemical staining of H. pylori on the surface of gastric mucosa is a valuable tool for identification of H. pylori infections.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
hCG is a protein secreted in large quantities by normal trophoblasts; the antibody detects cells and tumors of trophoblastic origin such as Choriocarcinoma. Large Cell Carcinoma and Adenocarcinoma of Lung demonstrate hCG positivity in 90% and 60% of cases respectively. 20% of Squamous Cell Lung Carcinomas are positive for hCG. hCG expression by nontrophoblastic tumors may indicate aggressive behavior since it has been observed that hCG may play a role in the host response to a given tumor.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Glypican-3 (GPC3) is a membrane-bound heparin sulfate proteoglycan known to participate in cell growth and differentiation. GPC3 expression has been observed in the majority of hepatocellular carcinomas (HCC), but is rarely expressed in non-neoplastic hepatic tissue, making it a useful marker for HCC. Additionally, this marker is expressed in many yolk sac tumors.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
1G12
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kandil DH, et al. Adv Anat Pathol. 2009; 16:125-9
References 2:
Coston WMP, et al. Am J Surg Pathol. 2008; 32:433-44
References 3:
Capurro M, et al. Gastroenterology. 2003; 125:89-97
References 4:
Zynger DL, et al. Am J Surg Pathol. 2006; 30:1570-5
Glycophorins A and B are major sialoglycoproteins expressed across the surface of the human erythrocyte membrane and contain the antigenic determinants that define the MNS blood group system.1 The high sialic acid content of glycophorin A contributes to the generation of a net negative surface charge across erythrocyte membranes that minimizes interactions between red blood cells and prevents their aggregation. Anti-glycophorin A has utility in identifying cells of the erythroid lineage.
Antibody Isotype:
IgG2b-k
Monosan Range:
MONOSAN Ready To Use
Clone:
GA-R2 & HIR2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Reid ME et al., Immunohematol 2009;25:95-101
References 2:
Olsen RJ et al. Arch Pathol Lab Med 2008; 132:462-75
Anti-Glucagon antibody detects glucagon-secreting cells and tumors such as glucagonomas. Studies show that approximately 80% of glucagonomas are malignant and these patients have a syndrome often initially recognized by dermatologists. Symptoms include necrolytic migratory erythema as well as diabetes, anemia, stomatitis, weight loss, frequent venous thromboses, and in some instances, diarrhea and psychiatric disturbances. The diagnosis may be readily confirmed by the demonstration of elevated plasma glucagon concentration.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Quesada I, et al. J Endocrinol. 2008; 199:5-19
References 2:
Gurlo T, et al. J Histotechnol. 2016; 39:8-16
References 3:
Wewer Albrechtsen NJ, et al. Biomark Med. 2016; 10:1141-51
Anti-GH is a useful marker in classification of pituitary tumors and the study of pituitary disease (acromegaly). It reacts with GH-producing cells. Growth hormone receptors have been found in various non-pituitary cells, including that from hepatocellular carcinoma and various benign and malignant cutaneous lesions.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Rezaei M, et al. J Res Med Sci. 2012; 17:681-5
References 2:
Al-Brahim NY, et al. J Clin Pathol. 2006; 59:1245-53
References 3:
Fukaya T, et al. Cancer. 1980; 45:1598-1603
References 4:
Kovacs K, et al. Virch Arch Pathol Anat. 1982; 395:59-68
The antibody detects astrocytes, Schwann cells, satellite cells, enteric glial cells, and some groups of ependymal cells. This marker is mainly used to distinguish neoplasms of astrocytic origin from other neoplasms in the central nervous system.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP672Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Jessen, KR, et al. J Neurosci 1983;3:2206-2218
References 2:
Regner A et al. J Neurotrauma. 2001 Aug;18(8):783-92
References 3:
Nagashima G et al. Clin Neurol Neurosurg. 2002 May;104(2):125-31
References 4:
Choi, BH, et al. Science 1984;223:407-409
References 5:
Viale, G, et al. Virchows Arch A Pathol Anat 1991;418:339-348
GCDFP-15 is a 15 kD glycoprotein which is localized in the apocrine metaplastic epithelium lining breast cysts and in apocrine glands in the axilla, vulva, eyelid, ear canal, and in salivary glands. GCDFP-15 positivity is seen in breast carcinomas. On the other hand, colorectal carcinomas, lung carcinoma, mesotheliomas rarely stain with this antibody. Because of its specificity for breast carcinoma, this antibody is often helpful in distinguishing metastasis of unknown primary.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
23A3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mazoujian G, et al. Am J Pathol. 1983; 110:105-12
References 2:
Liegl B, et al. Histopathology. 2007; 50:439-47
References 3:
Bhargava R, et al. Am J Clin Pathol. 2007; 127:103-13
References 4:
Tornos C, et al. Am J Surg Pathol. 2005; 29:1482-9
References 5:
Takeda Y, et al. Arch Pathol Lab Med. 2008; 132:239-43
Anti-Gastrin antibody gives positive staining of G-cells of human antral/pyloric mucosa and cells producing gastrin or a structural gastrin analogue as is seen in stomach; no staining of other cells or tissue types has been observed. This antibody may react with sulfated and non-sulfated forms of gastrin. The antibody cross-reacts with more than 50% of the present choleocystokinin octapeptide.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kasacka W, et al. Folia Morphol. 2012; 71:39-44.
References 2:
Hur K, et al. J Cancer Res Clin Oncol. 2006; 132:85-91
References 3:
Waldum et al. Frontiers in Endocrinology. 2017; 8:1-7
Galectin-3 is a 30-kD protein, a member of the beta-galactosidase-binding lectin family. Galectin-3 is associated with cell growth, adhesion, inflammation, mRNA processing, and apoptosis. Reportedly, Galectin-3 aberrant expression is related to malignant transformation and metastasis in carcinomas of the breast, colon and thyroid. Galectin-3 reactivity can be seen in the nucleus of neutrophils, vascular endothelium, carcinomas of the colon, breast, and thyroid. Galectin-3 may be useful in the differentiation of benign and malignant thyroid neoplasms. Galectin-3 may also be useful in the identification of certain liver disorders.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
9C4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Orlandi F, et al. Cancer Res. 1998; 58:3015-20
References 2:
Bartolazzi A, et al. Lancet. 2001; 357:1644-50
References 3:
Papotti M, et al. Eur J Endocrinol. 2002; 147: 515-21
Anti-FSH is a useful marker in classification of pituitary tumors and the study of pituitary disease. It reacts with FSH-producing cells (gonadotrophs).
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Baenziger JU, et al. Biochim Biophys Acta. 1988; 947:287-306
References 2:
Nussey SS, et al. BIOS Scientific Publishers Ltd; 2001 p. 217-79
References 3:
Uccella S, et al. Pituitary. 2000; 3:131-9
References 4:
Schmid M, et al. Pathol Res Pract. 2001; 197:663-9
Fascin is a 55-kd actin bundling protein involved in cell migration. Fascin is up-regulated in many human carcinomas and numerous studies have correlated Fascin over-expression with increased metastatic potential. Fascin is highly sensitive for staining Reed-Sternberg cells making it an excellent marker for classic Hodgkins lymphoma. It is uniformly negative in lymphoid cells, plasma cells and myeloid cells. MON 3279 is positive in dendritic cells. This marker may be helpful in distinguishing between Hodgkins disease and non-Hodgkins lymphoma in difficult cases.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
55-k2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Chu Pg Ann Diagn Pathol. 1999;3(2):104-33
References 2:
Pelosi G et al. Lung Cancer. 2003;42(2):203-13
References 3:
Goncharuk VN et al. J Cutan Pathol. 2002;29(7):430-8
References 4:
Kraus MD et al. Am J Dermatopathol. 2001;23(2):104-11
Factor XIIIa has been identified in platelets, megakaryocytes, and fibroblast-like mesenchymal or histiocytic cells in the placenta, uterus, and prostate, monocytes and macrophages and dermal dendritic cells. Anti-factor XIIIa has been found to be useful in differentiating between dermatofibroma (almost all cases +), dermatofibrosarcoma protuberans (-/+) and desmoplastic malignant melanoma. Anti-factor XIIIa positivity is also seen in capillary hemagioblastoma, hemangioendothelioma, hemangiopericytoma, xanthogranuloma, xanthoma, hepatocellular carcinoma, glomus tumor, and meningioma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
AC-1A1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Nemes Z, Hum Pathol 1992 Jul; 23(7):805-10
References 2:
Horenstein MG et al. Am J Surg Pathol. 2000 Jul;24(7):996-1003
References 3:
Kraus MD et al. Am J Dermatopathol. 2001 Apr;23(2):104-11
References 4:
Dehner LP. Am J Surg Pathol. 2003 May;27(5):579-93
References 5:
Deguchi M et al. Arch Dermatol Res. 2002 Oct;294(7):297-302
Anti-Factor VIII-Related Antigen antibody reacts with endothelial cells and neoplastic blood cells. This antibody has helped to establish the endothelial nature of some lesions of disputed histogenesis, e.g. Kaposis sarcoma and cardiac myxoma. Not all endothelial cells synthesize (or store) this molecule; therefore, it should not be surprising that not all tumors of endothelial differentiation (benign or malignant) react with this antigen.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Nichols GE, et al. Am J Clin Pathol. 1992; 97:770-5
References 2:
Falk S, et al. Am J Surg Pathol. 1993; 17:959-70
References 3:
Meis-Kindblom JM, et al. Am J Surg Pathol. 1998; 22:683-97
References 4:
Allison KH, et al. Am J Surg Pathol. 2004; 28:298-307
References 5:
Peyvandi F, et al. Blood Transfus. 2011; 9 Suppl 2:s3-8
The antibody 4 targets the 60 kDa latent membrane protein (LMP-1) encoded by the BNLF1 gene of the Epstein-Barr virus. There is cross-reactivity with Reed Sternberg cells of Hodgkins disease. The EpsteinBarr virus is an important as a cause of Infectious mononucleosis and has been associated with oral carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
CS1-4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Murray PG et al. J Pathol. 166: 1-5 (1992)
References 2:
Jarrett RF et al. Blood 78:1-10 (1991)
References 3:
- Pailesen G et al. Lancet. 337: 320-322 (1991)
References 4:
Silverberg GS et al Principles and Practice of Surgical Pathology and Cytopathology, 3rd edition. (1997)
Epithelial cell adhesion molecule (Ep-CAM) is a transmembrane glycoprotein localized on the membrane of cells in most epithelial tissues.1 Immunoreactivity with the antibody to Ep-CAM has been seen in the majority of epithelial neoplasms, whereas most non-epithelial neoplasms do not show Ep-CAM expression.2 Ep-CAM is not expressed in mesothelial cells, hepatocytes, and lymphocytes.1,2 In conjunction with other markers, Ep-CAM can be used as an aid in determining neoplasms of epithelial origin, such as distinguishing between lung adenocarcinoma and mesothelioma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MOC31
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Schnell U, et al. Biochim Biophys Acta. 2013; 1828:1989-2001
Epithelial cell adhesion molecule (Ep-CAM) is a transmembrane glycoprotein localized on the membrane of cells in most epithelial tissues. Immunoreactivity with the antibody to Ep-CAM has been seen in the majority of epithelial neoplasms, whereas most non-epithelial neoplasms do not show EpCAM expression. Ep-CAM is not expressed in mesothelial cells, hepatocytes, and lymphocytes. In conjunction with other markers, Ep-CAM can be used as an aid in determining neoplasms of epithelial origin, such as distinguishing between lung adenocarcinoma and mesothelioma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
Ber-EP4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Schnell U, et al. Biochim Biophys Acta. 2013; 1828:1989-2001
The antibody s a useful marker for staining many carcinomas. It stains normal and neoplastic cells from various tissues, including mammary epithelium, sweat glands and colorectal carcinoma. Hepatocellular carcinoma, adrenal carcinoma and embryonal carcinomas are consistently EMA negative, so keratin positivity with negative EMA favors one of these tumors. EMA is frequently positive in meningioma, which can be useful when distinguishing it from other intracranial neoplasms such as schwannomas.
Antibody Isotype:
IgG2a-k, IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
E29
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Dearnaly DP, et al. Lancet 1983; 1271-4
References 2:
Attanoos RL, et al. Histopathology. 2003; 43:231-8
E-cadherin is an adhesion protein that is expressed in cells of epithelial lineage. Anti-E-cadherin stains positively in glandular epithelium as well as adenocarcinomas of the lung, gastrointestinal tract and ovary. Another application involves the differentiation of ductal (which is membrane staining) vs. lobular cancer of the breast (which is cytoplasmic staining). It has also been shown to be positive in some thyroid carcinomas. A combination of E-cadherin and p120 catenin helps distinguish ductal carcinoma of the breast from lobular carcinoma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP700Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Han AC, et al. Hum Pathol. 1997; 28:641-5
References 2:
Simsir A, et al. Diagn Cytopathol. 1999; 20:125-30
References 3:
Abutaily AS, et al. J Clin Pathol. 2002; 55:662-8
References 4:
Acs G, et al. Am J Clin Pathol. 2001; 115:85-98
References 5:
Dabbs DJ, et al. Am J Surg Pathol. 2007; 31:427-37
Anti-desmin detects a protein that is expressed by cells of normal smooth, skeletal, and cardiac muscles. It has been suggested that desmin is primarily located at or near the periphery of Z lines in striated muscle fibrils. In smooth muscle, desmin interconnects cytoplasmic dense bodies with membrane bound dense plaques. Anti-desmin reacts with leiomyomas, leiomyosarcoma, rhabdomyomas, rhabdomyosarcoma, and perivascular cells of glomus tumors of the skin.This antibody is used to demonstrate the myogenic components/derivation of tumors. Desmin can also be present in myofibroblasts and be focally positive in desmoid fibromatosis.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
D33
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kouloumenta A, Journal of Biological Chemistry. 2007; 282:35211-21
References 2:
Altmannsberger M, et al. Am J Pathol 1985; 118:85-95
Podoplanin is a transmembrane mucoprotein (38 kD) recognized by the D2-40 monoclonal antibody. Podoplanin is selectively expressed in lymphatic endothelium as well as lymphangiomas, and Kaposi sarcomas. Podoplanin has also been shown to be expressed in epithelioid mesotheliomas and seminomas.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
D2-40
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ordóñez NG. Adv Anat Pathol. 2006; 13:83-8
References 2:
Niakosari F, et al. Arch Dermatol. 2005; 141:440-4
Human cytomegalovirus (CMV) is a ?-herpesvirus (human herpesvirus 5) that causes widespread persistent infection. CMV continues to be an important opportunistic pathogen in immunocompromised patients. It is estimated that 30% of transplant recipients experience CMV disease. The range of organ involvement in post-transplant CMV disease is wide; hepatitis occurs in 40% of liver transplant recipients, and pneumonitis is more frequently seen in heart and heart-lung transplant patients. Other organs that are commonly affected are the gastrointestinal tract and the peripheral and central nervous systems. Histologic diagnosis of CMV in fixed tissues usually rests on identifying characteristic cytopathic effects including intranuclear inclusions, cytoplasmic inclusions, or both, especially in the endothelial cells. However, histologic examination lacks sensitivity, and in some cases atypical cytopathic features can be confused with reactive or degenerative changes.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
8B1.2,1G5.2&2D4.2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Drummer, JS et al. J Infect Dis 1985; 152:1182-1191
References 2:
Cote, L et al. J Clin Microbiol 1993; 31:2066-2069
The antibody is the broad-spectrum keratin antibody cocktail. It is composed of mouse monoclonal antibody AE1 that recognizes the acidic type I keratins 10, 14, 15, 16, 19, and AE3 that reacts with the basic type II keratins 1, 2, 3, 4, 5, 6, 7, and 8. Both clones were generated using epidermal keratin as immunogen. This antibody detects carcinomas of different organ origin, but is most frequently negative in hepatocellular carcinoma, chromophobe RCC, adrenol cortical carcinoma, some clear cell renal cell carcinomas, and renal oncocytoma. This antibody cocktail can cross-react with other intermediate filaments, such as glial fibrillary acidic protein, giving a false-positive staining in glial tumors.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
AE1/AE3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Anti-cytokeratin, low molecular weight (AE1) antibody labels acidic keratins K10, K14, K15, K16, and K19. Anti-cytokeratin (AE1) reactivity is seen in most normal and neoplastic cells of epithelial origin. Anti-cytokeratin (AE1) is a useful immunohistochemical reagent for distinguishing between poorly differentiated carcinomas and non-epithelial neoplasms.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
AE1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Tyler, CR. Arch Pathol Lab Med. 1978; 102:113-21
References 2:
Weiss RA, et al. J Cell Biol. 1984; 98:1397-406
References 3:
Lopez-Beltran A, et al. Virchows Arch. 2001; 438:552-7
References 4:
Kitazawa R, et al. Virchows Arch. 1999; 435:137-42
References 5:
Judkins AR, et al. Am J Clin Pathol. 1998; 110:641-6
Anti-cytokeratin, high molecular weight (AE3) is capable of recognizing all basic keratins; therefore, it is a broadly reactive antibody staining most epithelia and their neoplasms. Members of the acidic and basic subfamilies are found together in pairs. Since each epithelium contains at least one acidic and one basic keratin, this antibody is used to observe the distribution of keratin-containing cells in normal epithelia and to identify neoplasms derived from such epithelium.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
AE3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Tyler CR.Arch Pathol Lab Med. 1978; 102:113-21
References 2:
Weiss RA, et al. J Cell Biol. 1984; 98:1397-406
References 3:
Lopez-Beltrán A, et al. Virchows Arch. 2001; 438:552-7
The antibody is an antibody to high molecular weight cytokeratin that reacts with all squamous and ductal epithelium and stains carcinomas. This antibody recognizes cytokeratins 1,5,10, and 14 that are found in complex epithelia. Anti-Cytokeratin, 34betaE12 shows no reactivity with hepatocytes, pancreatic acinar cells, proximal renal tubules, or endometrial glands; there has been no reactivity with cells derived from simple epithelia. Mesenchymal tumors, lymphomas, melanomas, and neural tumors are unreactive with this antibody with some exceptions. Anti-Cytokeratin, 34betaE12 does label myoepithelial cells and has been shown to be useful in distinguishing prostatic adenocarcinoma from hyperplasia of the prostate. This antibody has also been useful in separating benign from malignant intraductal breast proliferations.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
34betaE12
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gown AM, et al. Am J Pathol. 1984; 114:309
References 2:
OMalley FP, et al. Virch Arch A. 1990; 417:191-6
References 3:
Wojno KJ, et al. Am J Surg Pathol. 1995; 19:251-60
References 4:
Moinfar F, et al. Am J Surg Pathol. 1999; 23:1048-58
Cytokeratin 20 is a 46-kDa intermediate filament protein which reacts primarily with gastric and intestinal epithelium, urothelium, and Merkel cells.1-4 Anti-Cytokeratin 20 is useful in the identification of specific types of these epithelial cells under normal hyperplastic and neoplastic conditions.3 Cytokeratin 20 has been detected in adenocarcinomas of the colon, stomach and biliary tract.5-7 Merkel cell carcinomas have shown reactivity. In contrast, carcinomas of the breast are generally non-reactive.
Antibody Isotype:
IgG2a-k
Monosan Range:
MONOSAN Ready To Use
Clone:
Ks20.8
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Anti-Cytokeratin 19 reacts with a wide variety of epithelium and epithelial malignancies including Adenocarcinomas of the Colon, stomach, pancreas, biliary tract, liver and breast. Perhaps the most useful application is the identification of Thyroid carcinoma of the Papillary type, although Follicular Carcinoma is also labeled by this antibody approximately 50-60% of the time.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
A53-B/A2.26
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Cerilli LA, et al. Am J Clin Pathol 2002;118:186-193
References 2:
Jain R, et al. Appl Immunohistochem Mol Morphol. 2010; 18:9-15
Cytokeratin 14 is a member of the Type I family of cytokeratins and is generally expressed in the basal cell layer of squamous epithelium. The cytokeratin 14 protein forms a heterotetramer with homodimers of cytokeratin 5 to contribute to the structural integrity of the intracellular cytoskeletal network. Anticytokeratin 14 has immunohistochemical utility as an aid in distinguishing squamous cell carcinomas from other tumors of epithelial origin.
Antibody Isotype:
IgG3
Monosan Range:
MONOSAN Ready To Use
Clone:
LL002
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Reis-Filho JS et al. Appl Immunohistochem Mol Morphol; 11(1):1-8 (2003)
Cytokeratins 8 &18 (CK 8 & 18) are expressed in most simple epithelia (e.g. thyroid, breast, gastrointestinal tract, and respiratory tract). Anti-CK 8 & 18 have been reported to stain most adenocarcinomas and squamous cell carcinomas, but not some well-differentiated squamous cell carcinomas. Cytokeratin 8 & 18 have been reported to be useful markers for identifying Paget cells, colorectal carcinoma metastases,5 and gastric cancer micro metastases.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
B22.1&B23.1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Angus B, et al. J Pathol. 1987; 155:377-84
References 2:
Corson, JM. Pathol Annu. 1986; 21:47-81
References 3:
Moll R, et al. Histochem Cell Biol. 2008; 129:705-33
Cytokeratin 8, a member of the Type II family of cytokeratins, is typically expressed in simple epithelium. The dimerization of cytokeratin 8 with cytokeratin 18 (labeled by 35betaH11) in the cytoplasm of simple epithelial cells allows for the formation of an intermediate filament cytoskeletal framework. This structure plays a role in the maintenance of cellular structural integrity and also functions in promoting signal transduction and cellular differentiation processes. Additionally, the presence of cytokeratin 8 has been detected in neoplastic epithelia, including glandular epithelium that can be found in prostate carcinoma. Positive immunoreactivity with anti-cytokeratin 8 is a useful indicator for the identification of normal and neoplastic epithelial tissues.
Antibody Isotype:
IgM
Monosan Range:
MONOSAN Ready To Use
Clone:
35betaH11
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Battifora, H. Am J Surg Pathol 1988;12:24
References 2:
Gown, AM, et al. Am J Clin Pathol 1985;84:413
References 3:
Ljung G, et al. Prostate. 1997; 31:91-7
References 4:
Murata T, et al. Pathol Res Pract. 1993; 189:888-93
References 5:
Moll R, et al. Histochem Cell Biol. 2008; 129:705-33
Anti-cytokeratin 7 reacts with proteins that are found in most ductal, glandular, transitional, and biliary duct epithelial cells. Cytokeratin 7 (CK7) labeling can help distinguish between lung,breast carcinomas, and urothelial carcinomas that typically stain positive, and colon and prostate carcinomas that typically lack CK7 expression.CK 7 is a common marker of primary lung adenocarcinomas (almost all cases) with a lower specificity since it is also observed in other primary lung carcinomas and non-pulmonary carcinomas.1 Anti-cytokeratin 7 has also been useful in the differential diagnosis of ovarian neoplasms.This antibody does not recognize intermediate filament proteins.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
OV-TL 12/30
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Jerome MV, et al. Histopathology. 2004; 45:125-34
References 2:
Murray SK, et al. Am J Surg Pathol. 2004; 28:1154-62
References 3:
Ramalingam P, et al. Ann Diagn Pathol. 2003; 7:112-9
References 4:
McCluggage WG, et al. Histopathology. 2005; 47:231-247
References 5:
Roy S, et al. Arch Pathol Lab Med. 2011; 135:1601-5
Anti-CK 5/6 positivity is seen in nearly 100% of malignant mesotheliomas and in nearly 0% of lung adenocarcinomas. Anti-CK 5/6 positivity can be seen in undifferentiated large cell carcinoma as well as squamous carcinoma, and has been useful in recognizing spindle cell squamous cell carcinoma of the skin. Less than 10% of carcinomas of the breast, colon, and prostate stain positively for this marker. Anti-CK 5/6 has also been used successfully as a myoepithelial cell marker in the prostate and breast to determine malignancy.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
D5/16B4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ordonez NG. Am J Surg Pathol 1998; 22(10):1215-1221
References 2:
Abarahams NA et al. Am J Clin Pathol. 2003 Sep;120(3):368-76
References 3:
Reis-Filho JS et al. Virchows Arch. 2003 Aug;443(2):122-32
References 4:
Lin L et al. J Cutan Pathol.2003 Feb;30(2):114-7
References 5:
Otterbach F et al. Histopathology. 2000 Sep;3793):232-40
Cytokeratin 5 is an intermediate filament protein of 58 kD molecular weight within the cytokeratin family. It is a type II (basic) cytokeratin. Antibodies to this protein identify basal cells of squamous and glandular epithelia, myoepithelia, and mesothelium. Anti-cytokeratin 5 has been reported useful in the differential diagnosis of metastatic carcinoma in the pleura versus epithelioid mesothelioma. Epithelioid mesotheliomas are strongly positive in almost all cases, but a minority of pulmonary adenocarcinomas will show focal immunoreactivity. Almost all squamous cell carcinomas, half of transitional carcinomas, and many undifferentiated large cell carcinomas immunostain with anti-CK 5. Anti-CK 5, along with antip63, affords a high sensitivity and specificity for squamous differentiation. Myoepithelial cells of the breast, glandular epithelia, and basal cells of the prostate are labeled with anti-CK. This antibody, along with anti-CK 14, has found application in identifying basal-like breast carcinoma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP1601Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Ordonez NG. Human Pathology. 2007; 38:116
References 2:
Kargi A, et al. Appl Immunohistochem Mol Morphol.; 15:415420 (2007)
Cyclooxygenase 2 (COX-2) is an essential enzyme involved not only in the mediation of inflammation but also carcinogenesis. Increased expression of COX-2 has been shown in carcinomas of many organ systems including stomach, colorectum, breast and lung.
Antibody Isotype:
IgG-1
Monosan Range:
MONOSAN Ready To Use
Clone:
SP21
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Collagen Type IV is the major component of the basal lamina so antibodies to this molecule confirm its presence and reveal the morphological appearance of the structure. Normal tissue stains with this antibody in a fashion consistent with the sites of mesenchymal elements and epithelial basal laminae. Anti-Collagen IV can also be useful in the classification of soft tissue tumors; schwannomas, leiomyomas. Their well differentiated, malignant counterparts usually immunoreact with this antibody. The vascular nature of neoplasms, hemangiopericytoma, angiosarcoma and epithelioid hemangioendothelioma can be revealed by this antibody with greater reliability than non-specific stains (e.g. silver reticulum).
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
CIV22
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gould, VE, et al., Pathol Annul 1976;11:353-386
References 2:
McArdle, JP, et al., Int J Cancer 1984;34:633-638
References 3:
De Iorio P et al. Anticancer Res. 2001;21(2B):1395-9
References 4:
Maatta M et al. J Histochem Cytochem. 2001 Jun;49(6):711-26
References 5:
Schmehl K et al. Int J Colorectal Dis. 2000 Feb;15(1):39-48
Immunohistochemical methods have localized chromogranin in a wide variety of endocrine tissues including the pituitary, pancreas, thyroid, and parathyroid. Neuroendocrine cells exhibit a fine granular immunoreactivity to chromogranin. It is generally accepted that the co-expression of certain keratins and chromogranin mean neuroendocrine lineage. The presence of strong chromogranin staining and absence of keratin staining should raise the possibility of paraganglioma. The co-expression of chromogranin and NSE is typical of neuroendocrine neoplasms.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
LK2H10
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Wilson, BS, et al., Am J Pathol; 115:458-468 (1984)
References 2:
Lyda MH, Weiss LM. Hum Pathol. 31(8):980-7 (2000)
References 3:
ontochristopoulous GJ et al. Dermatology.; 201(2):123-6 (2000)
References 4:
Qvigstad G et al. Histochem J.; 32(9):551-6 (2000)
Anti-CEA specifies a group of proteins in the Carcinoembryonic Antigen (CEA) family of proteins which are present in the epithelia of various types and tumors (both benign and malignant) derived from such epithelia. Such tissues are represented by the epithelia of colon, bronchus, alveoli, breast, pancreas, biliary tract, superficial layer and parietal layers of the stomach. Predominately biliary canaliculi are labelled in the liver and this factor is useful in the diagnosis of hepatocelluar carcinoma. Anti-CEA has been quite useful in differentiating adenocarcinoma of the lung vs. mesothelioma. Associated products: CK 5/6, Calretinin, WT-1, E-Cadherin, TTF-1, TAG-72, EMA, CK 20
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Shield PW, et al. Am J Clin Pathol. 1996; 105:157-62
References 2:
Sheahan K, et al. Am J Clin Pathol. 1990; 94:157-64
Anti-CEA is employed as a tool to assist in the distinction between adenocarcinoma and mesotheliomas, along with other markers such as calretinin, CK 5/6, D2-40, HBME-1, Napsin A, MOC31, and Ber-EP4. Anti-CEA positivity is seen in adenocarcinomas from the lung and colon, as well.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
CEA31
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Go, VLW, et al., Cancer 1976;37:562-566
References 2:
Delellis, RA, et al., Am J Clin Pathol 1978;50:587-594
References 3:
Abutaily AS et al. J Clin Pathol. 2002 Sep;55(9):662-8
References 4:
Bhatnagar J et al. Anticancer Res. 2002;22(3):1849-57
References 5:
Carella R. et al. Am J Surg Pathol. 2001 Jan;25(1):43-50
CD138, Syndecan 1, is expressed in the late stages of B-cell differentiation with progression towards plasma cells. It can be used to differentiate lymphoplasmacytic lymphoma from marginal zone lymphoma. ALK+ large B-cell lyphoma (LBCL) usually strongly expresses CD138 whereas lineageassociated markers such as anti-CD20 and anti-CD79a do not stain ALK+LBCL. Anti-CD138 is immunoreactive with HHV8-associated primary effusion lymphoma even though the lymphoma cells lack the expression of B-cell markers. Anti-CD138 is a good marker to identify and enumerate plasma cells, benign, reactive, or malignant, in bone marrow biopsy specimens.4,6 CD138 is also expressed in epithelial cells.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
B-A38
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Chilosi M, et al. Mod Pathol. 1999; 12:1101-6
References 2:
Sebestyén A, et al. Br J Haematol. 1999; 104:412-9
References 3:
Bayer-Garner IB, et al. Mod Pathol. 2001; 14:1052-8
References 4:
OConnell FP, et al. Am J Clin Pathol. 2004; 121:254-63.
References 5:
Colomo L, et al. Am J Surg Pathol. 2004; 28:736-47
CD163, also known as scavenger receptor cysteine-rich type 1 protein M130, is an acute phase-regulated and signal-inducing transmembrane protein, found exclusively on cells of monocytic origin. CD163 plays a critical role in macrophage clearance and endocytosis of hemoglobin/haptoglobin complexes. Therefore, CD163 contributes to the anti-inflammatory response and protects tissues from oxidative and inflammatory hemoglobin. Anti-CD163 labels cells of monocytic-macrophage lineage, with expression in bone marrow3 and histiocytic neoplasms. Solubilized in plasma, CD163 functions as an anti-inflammatory signal and has many roles in disease processes that range from autoimmune conditions such as rheumatoid arthritis to atherosclerosis.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-26
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Buechler C, et al. J Leukoc Biol. 67:97-103 (2000)
References 2:
Kristiansen M, et al. Nature. 409:198-201 (2001)
References 3:
Etzerodt A. et al. Antioxid Redox Signal. 18: 2352-63 (2013)
CD117, c-kit is a tyrosine kinase receptor found on interstitial cells of Cajal, germ cells, bone marrow stem cells, melanocytes, breast epithelium and mast cells. This receptor is found on a wide variety of tumor cells (follicular and papillary carcinoma of thyroid, adenocarcinomas from endometrium, lung, ovary, pancreas, breast; malignant melanoma, endodermal sinus tumor, and small cell carcinoma) but has been particularly useful in differentiating gastrointestinal stromal tumors from Kaposis sarcoma, and tumors of smooth muscle origin.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
YR145
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Sircar K, et al. AM J Surg Pathol 23(4):377-389,1999
References 2:
Miettinen M et al. Am J Surg Pathol 24(2):211-222, 2000
References 3:
Miettinen M. et al. Am J Surg Pathol 23(9): 1109-1118
The antibody is a B-cell marker that is generally used to complement CD20. This antibody will stain many of the same lymphomas as CD20, but also is more likely to stain precursor B-lymphoid leukemias than CD20. Anti-CD79a also stains more cases of plasma cell myeloma and occasionally some types of endothelial cells as well. Anti-CD79a will stain many cases of acute promyelocytic leukemia (FAB-M3), but only rarely stains other types of myeloid leukemia.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
JCB117
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mason DY, et al., Eur J Immun; 22:2753-2756 (1992)
References 2:
Lin BT, Weiss LM. Hum Pathol.; 28(9):1083-90 (1997)
References 3:
Pilozzi E et al. J Pathol.; 186(2):140-3 (1998)
References 4:
Kurtin PJ et al. Am J Clin Pathol.; 112(3):319-29 (1999)
References 5:
Blakolmer K et al. Mod Pathol.; 13(70:766-72 (2000)
The antibody marks cells of monocyte/macrophage lineage. This antibody is capable of staining monocytes, Kupffer cells, osteoclasts, granulocytes and their precursors; lymphomas are negative or show few granules. This antibody may be useful for the identification of myelomonocytic and histiocytic tumors. Since this detects a formalin-resistant epitope that may be associated with lysosomal granules, other lysosome-rich cells may also stain.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
Kp-1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
CD63 is a 53kDa lysosomal membrane protein in the family of tetraspan moieties, and characterized as an activation dependent platelet surface antigen. Anti-CD63 reactivity is seen in the cytoplasm of many cell types including lymphoid, myeloid, endothelial cells, and the majority of malignant melanomas. Anti-CD63 is a useful immunohistochemical marker for the identification of malignant melanoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
NKI/C3
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Azorsa DO, et al. Blood. 1991; 78:280-4
References 2:
Barrio MM, et al. Hybridoma. 1998; 17:355-64
References 3:
Demetrick DJ,et al. J Natl Cancer Inst. 1992; 84:422-9
CD61 also known as integrin beta chain beta 3 (ITGB3) is an integrin cell-surface protein associated with cellular adhesion and cell-surface mediated signaling. Immunohistochemical staining for CD61 can be useful in evaluating normal and abnormal megakaryocytes, which can aide in the identification of some hematopoietic malignancies. Anti-CD61 reactivity is also seen in platelets, osteoclasts and macrophages.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
2f2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Duperray A et al. Blood. 1989 Oct; 74(5):1603-11
References 2:
Goldman BI et al. Modern Pathology 14:589-594 (2001)
References 3:
Thiele J et al. Virchows Archiv B Cell Pathol (1990) 58:295-302
CD57, also known as HNK-1 (human natural killer-1), is a cell surface carbohydrate epitope expressed on terminally differentiated T-cells and subsets of natural killer (NK) cells.1 It has also been identified on cells of neural crest origin.2 Anti-CD57 is often used to visualize the non-neoplastic bystander T-cells that may form rosettes around the neoplastic lymphocyte-predominant (LP) cells in nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL).3
Antibody Isotype:
IgM-k
Monosan Range:
MONOSAN Ready To Use
Clone:
NK1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kared H, et al. Cancer Immunol Immunother. 2016; 65:441-52
References 2:
Nielsen CM, et al. Front Immunol. 2013; 4:422
References 3:
Sattarzadeh A, et al. Exp Hematol Oncol. 2015; 4:27
CD56, also known as neural cell adhesion molecule (NCAM), is a calcium-independent homophilic binding protein that belongs to a group of cell adhesion molecules including cadherins, selectins, and integrins. CD56 is involved in cellcell adhesion of neural cells during embryogenesis and is expressed on most neuroectodermally derived tissues. In normal tissue, anti-CD56 labels neurons, glia, schwann cells, NK (natural killer) cells, and a subset of T-cells.3 CD56 expression can be seen in most NK cell neoplasms, certain subtypes of T-cell lymphoma and in some plasma cell neoplasms. well
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
123C3.D5
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Langdon, SP, et al. Cancer Research 1988;48(21):6161-6165
References 2:
Moolenaar, CE, et al. Cancer Research 1990;50(4):1102-1106
References 3:
Sumi M et al. Leuk Lymphoma. 2003 Jan; 44(1): 201-4
References 4:
Kibbelaar, RE, et al. Euro J of Cancer 1991;27(4):431-435
References 5:
Michalides, R, et al. International J of Cancer Sup 1994;8:34-37
Anti-CD45RO labels an isoform of the CD45 antigen also known as leukocyte common antigen. Anti-CD45RO reacts with thymocytes, mature activated T-cells, and a subpopulation of resting T-cells while showing no reactivity with B-cells, making this antibody helpful in identifying T-cell neoplasms.
Antibody Isotype:
IgG2a-k
Monosan Range:
MONOSAN Ready To Use
Clone:
UCHL-1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Anti-CD45 (anti-leukocyte common antigen) is routinely used to aid the differential diagnosis of undifferentiated neoplasms, whenever malignant lymphoma is suspected by the morphological or clinical data. It is a highly specific antibody; therefore a positive result is highly indicative of hematolymphoid origin. Certain types of hematolymphoid neoplasms may lack CD45 (Hodgkin lymphoma, some T-cell lymphomas, and some leukemias) so its absence does not rule out a hematolymphoid tumor. This antibody is expressed almost exclusively by cells of hematopoietic lineage and is present in most benign and malignant lymphocytes as well as plasma cell precursors.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
2B11 & PD7/26
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mason, DY, Am Pathol 1987;128:1-4
References 2:
Hall PA, Histopathology 1988;13:149-160
References 3:
Kurtin, PJ, Hum Path 1985;16:353-365
References 4:
Maluf HM et al. Mod Pathol. 1995 Feb; 8(2): 155-9
References 5:
Caballero T et al. J Clin Pathol. 1995 Aug;48(8): 743-8
The CD44 family of glycoproteins exists in a number of variant isoforms, the most common being the standard 85-95 kD or hematopoietic variant (CD44s) that is found in mesodermal cells such as hematopoietic, fibroblastic, and glial cells, as well as in some carcinoma cell lines. Higher molecular weight isoforms have been described in epithelial cells (CD44v) and are thought to function in intercellular adhesion and stromal binding. While many human tumors express CD44, a positive correlation between CD44v expression and tumor dedifferentiation has been demonstrated. MON 3237 may be useful in discrimination of urothelial carcinoma in-situ from non-neoplastic changes in the urothelium.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-13
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Hudson D, et al. Int. J. Cancer. 1996; 66:457-63
References 2:
East JA, et al. Eur J Cancer. 1993; 29A:1921-2
References 3:
Gadalla HA, et al. BJU Int. 2004; 93:151-5
References 4:
McKenney JK, et al. Am J Surg Pathol. 2001; 25:1074-8
References 5:
Lopez-Beltran A, et al. Anal Quant Cytopathol Histpathol. 2013; 35:121-9
CD43 is a transmembrane protein involved in immune function and T-cell activation. AntiCD43 reactivity is seen in T lymphocytes, monocytes, and granulocytes. No reactivity has been observed in normal or reactive B-cells. Reportedly, anti-CD43 reactivity is seen in the majority of T-cell lymphomas and some low grade B-cell lymphomas. Therefore, anti-CD43 is a useful immunohistochemical marker for the identification of T-cell lymphomas and some low grade B-cell lymphomas
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
MT1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Leong AS-Y, et al. 2nd edition. Grenwich Medical Media. London. 2003
References 2:
Dabbs DJ. Diagnositc Immunohistochemistry. Third Edition. Saunders. 2006
CD34 is a cell surface glycophosphoprotein expressed on human hematopoietic progenitor cells and can be used for identifying blast cells. CD34 is a marker for vascular endothelial cells and has been shown in literature to be highly sensitive for angiosarcomas and Kaposi's sarcomas. In addition, CD34 is expressed in soft tissue tumors such as gastrointestinal stromal tumors (GIST).
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
QBEnd/10
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Civin, CL, et al., London Academic Press 1989:818-825
References 2:
Fina, L et al., Blood 1990;75:2417-2426
References 3:
Torlakovic G et al. Arch Pathol Lab Med. 2002 Jul;126(7):823-8
References 4:
Salizzoni M et al. Transplantation 2003 Sep 15;76(5):844-8
References 5:
Fanburg-Smith JC et al. Mod Pathol. 2003 Mar;16(3):263-71
CD31 has cytoplasmic, membranous expression in non-neoplastic and neoplastic vascular endothelial cells.1 It has been used as a tool to identify the vascular origin of neoplasms such as angiosarcomas, Kaposi sarcomas and epithelioid hemangioendothelioma. Immunohistochemical study with CD31 has also been shown useful to detect areas of tumor lymphovascular invasion. Additionally, detection of weak diffuse cytoplasmic CD31 immunoreactivity has been seen in cases of various carcinomas with occasional membranous staining in ductal carcinomas of the breast as well as in intratumoral macrophages.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
JC70
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Parums DV, et al.. J Clin Pathol. 1990; 43:752-7
References 2:
Attanoos RL, et al. Thorax. 2000; 55:860-3
References 3:
Alexander-Sefre F, et al. J Clin Pathol. 2003; 56:786-8
The antibody detects a formalin-resistant epitope that is expressed by Reed-Sternberg cells in classic Hodgkin lymphoma, the majority of anaplastic large cell lymphomas, primary cutaneous CD30 positive T-cell lymphoproliferative disorders and in embryonal carcinomas. Occasionally diffuse large B-cell lymphoma stains with this antibody. This antibody also stains plasma cells in paraffin-embedded tissue as well as reactive immunoblasts. The staining pattern of anti-CD30 in lymphoma and embryonal carcinoma is different, with the former being membranous and exhibiting Golgi zone accentuation in location, and the latter being membranous only.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
Ber-H2
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Schwarting R, et al., Blood 1989 (74):1678-1689
References 2:
Fonatsch C, et al., Genomics 1992 (14):825-826
References 3:
George DH et al. Am J Surg Pathol. 2003 Apr;27(4): 487-93
References 4:
Hedvat CV et al. Hum Pathol. 2002 Oct;33(10): 968-74
CD25, Interleukin-2 receptor alpha chain, is the alpha subunit of the cell surface receptor which regulates regulatory T-cells. CD25 has been detected in various hematological malignancies including adult T-cell leukemia/lymphoma, and hairy cell leukemia. MON 3230 has also been useful in identifying mast cells in skin biopsies in the setting of Urticaria Pigmentosa, which is predictive of Systemic Mastocytosis.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN Ready To Use
Clone:
4C9
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Hahn HP, et al. Am J Surg Pathol. 2007 Nov;31(11):1669-76
References 2:
Hollmann TJ, et al. Am J Surg Pathol. 2008 Jan;32(1):139-45
References 3:
Létourneau S, et al. Clin Immunol. 2009; 123:758-62
References 4:
De Totero D, et al.. Leuk Lymphoma. 1994; 104:412-9
References 5:
Qayyum S, et al. Archives of Pathology & Lab Med. 2014; 138:282-6
CD23 antigen is a 45-60 kDa membrane glycoprotein identified as a low affinity receptor for IgE production as well as a receptor for lymphocyte growth factor. CD23 is found in some mature B-cell lymphomas and in Reed-Sternberg cells in Hodgkin disease. Follicular dendritic cells and some activated B-cells within germinal centers express CD23 in high density and mantle zone B-cells are stained. The majority of chronic lymphocytic leukemias/small lymphocytic lymphomas are anti-CD23 positive, whereas mantle cell lymphomas are generally negative, so this marker is useful when applied with other markers to separate the small cell lymphomas.1,3 Precursor B and T lymphomas, myeloid neoplasms, and mature T-cell lymphomas are CD23 negative and other small cell lymphomas are occasionally positive. Anti-CD23 is expressed in activated mature B-cells expressing IgM or IgD, monocytes/macrophages, follicular dendritic cells, T-cell subsets, eosinophils, Langerhans cells and small lymphocytic lymphoma/chronic lymphocytic leukemia.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
1B12
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kaiserlian, D, et al., Immunology 1993;80:90-95
References 2:
Aubry, JP et al., Oxford Univ Press- Oxford, NY, Tokyo 1987:417-419
References 3:
Pallesen G, Oxford Univ Press-Oxford, NY, Tokyo 1987:383-386
References 4:
Pezzella F et al. Br j Haematol. 2000 Feb;108(2): 369-76
References 5:
Kurtin PJ et al. Am J Clin Pathol. 1999 Sep;112(3): 319-29
CD21 (also known as complement receptor 2 (CR2), C3d receptor, or EBV receptor) is a 140 kDa membrane protein on B-lymphocytes to which the Epstein-Barr virus (EBV) binds during infection of these cells. The antigen is absent on T-lymphocytes, monocytes, and granulocytes. MON 3027 is useful in the identification of follicular dendritic cell matrix found in normal lymph node and tonsillar tissue. This antibody also labels follicular dendritic cell sarcomas. Anti-CD21 is valuable in differentiating follicular lymphoma with marginal zone differentiation from marginal zone lymphoma with follicular involvement. It also plays a role in distinguishing among nodular lymphocyte predominant Hodgkin lymphoma, lymphocyte-rich classic Hodgkin lymphoma, and T-cell/histiocyte-rich B-cell lymphoma in combination with other B-cell and T-cell markers.6 Anti-CD21 is also useful in identifying abnormal follicular dendritic cell pattern in angioimmunoblastic T-cell lymphoma and follicular T-cell lymphoma.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN Ready To Use
Clone:
2G9
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Dillon KM et al. J Clin Pathol.; 55(10):791-4 (2002)
References 2:
Cheuk W, et al. Am J Surg Pathol.; 25:721-31 (2001)
References 3:
Pileri SA, et al.Histopathology.; 41:1-29 (2002)
References 4:
Maeda K, et al. J Histochem Cytochem.; 50:1475-1486 (2002)
CD20 is a transmembrane protein in late B-cell precursors and mature B-cells that plays a role in regulating proliferation and differentiation. CD20 expression is lost at the plasma cell stage of differentiation. MON 3226 (pan B-cell) has rarely been detected in T-cell malignancies, and is a dependable marker of B-cell lymphomas such as DLBCL. CD20 expression is present in some thymomas. It does not cross-react with non-hematopoietic neoplasms.
Antibody Isotype:
IgG2a-k
Monosan Range:
MONOSAN Ready To Use
Clone:
L26
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
CD19 is a glycoprotein on the surface of mature B cells, it works in conjunction with receptors and proteins to regulate B-cell signaling. CD19 is present in both normal and malignant B cells, and hence being valuable for the identification of B-cell neoplasms such as diffuse large B-cell lymphoma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-36
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Steinmetz OM, et al. Transplantation. 2007 15;84(7):842-50
References 2:
Teng YK, et al. Arthritis Rheum. 2007 ;56(12):3909-18
CD15 is a carbohydrate antigen with the common trisaccharide structure 3-fucosyl-N-acetyllactosamine, also known as Lewis x (Lex) or stage-specific embryonic antigen 1 (SSEA-1).1-3 CD15 is expressed in myeloid cells and mediates neutrophil adhesion to dendritic cells.2-3 CD15 is also expressed in Reed-Sternberg cells and is thus a useful marker for identifying Hodgkins lymphoma.
Antibody Isotype:
IgM
Monosan Range:
MONOSAN Ready To Use
Clone:
MMA
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Pellegrini W, et al. Haematologica. 2007; 92:708-9
Common acute lymphoblastic leukemia antigen (CALLA / CD10) is a useful marker for the characterization of childhood leukemia and B cell lymphomas. This antibody reacts with antigen of lymphoblastic, Burkitts, and follicular lymphomas; and chronic myelocytic leukemia. Also, Anti-CD10 detects the antigen of glomerular epithelial cells and the brush border of the proximal tubules; this characteristic may be helpful in interpreting renal ontogenesis in conjunction with other markers. Other non-lymphoid cells that are reactive with CD10 are breast myoepithelial cells, bile canaliculi, neutrophils and small population of bone marrow cells, fetal small intestine epithelium, and normal fibroblasts.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
56C6
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Maguer-Satta V, et al.. Stem Cells. 2011; 29:389-96
The CD8 (cluster of differentiation 8) antigen is a cell surface glycoprotein made up of two subunits alpha and beta.1 Anti-CD8 is a T-cell marker for the detection of cytotoxic/suppressor lymphocytes. CD8 is also detected on NK cells, some thymocytes, some null cells and bone marrow cells. This antibody, along with other markers, can be used to distinguish between reactive and neoplastic Tcells.3 CD8 expression has been found to be negative in Mycosis Fungoides. Rarely does anti-CD8 label non-hematolymphoid neoplasms.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
C8/144B
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Rossi, ML, Sanchez, FC, et al., J Clin Path 1988;41:314-319
References 2:
Stein, H, Lennart, K, et al., Adv Cancer Res 1984;42:67-147
References 3:
Phan-Dinh-Tuy, F, Niaudet, P, et al., Mol Immun 1982;19:1649-1654
CD7 antigen is a 40-kDa cell surface glycoprotein that is a member of the immunoglobulin gene superfamily. While its precise function is not known, it is suggested that CD7 plays a role in T-cell interactions as it is one of the earliest T-cell lineage associated antigens expressed during T-cell ontogeny. CD7 is expressed in thymocytes, mature peripheral T-cells, natural killer cells, and lymphoid and myeloid progenitors. CD7 is the most consistently expressed T cell antigen in lymphoblastic lymphomas and leukemias, and is therefore a useful marker in the identification of such neoplastic proliferations. In mature post-thymic T cell neoplasms, it is the most common pan-T antigen to be aberrantly absent and its absence in a T cell population is a useful pointer to a neoplastic conversion.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
MRQ-56
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Hodak E, et al. J Am Acad Derma¬tol. 2006 Aug;55(2):276-84
References 2:
Stillwell R, et al. Immunol Res. 2001; 24:31-52
References 3:
Schanberg LE, et al. Proc Natl Acad Sci USA. 1991; 88:603-7
Anti-CD5 is a pan T-cell marker that also reacts with a range of neoplastic B-cells. CD5 expression is useful in distinguishing mature T-cell neoplasms and differentiating among mature small lymphoid cell malignancies. Anti-CD5 does not react with granulocytes or monocytes.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
4C7
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Jones NH, et al., Nature 1986;323: 346-349
References 2:
Tan SH et al. Br J Dermatol. 2003 Sep;149(3): 542-53
References 3:
Chang CC et al. Mod Pathol. 2002 Oct;15(10): 1051-7
References 4:
Hatano B et al. Pathol Int. 2002 May-Jun;52(5-6): 400-5
References 5:
West RB et al. Am J Clin Pathol. 2002 Apr;117(4): 636-43
Anti-CD3 antibody has been considered the best all around T-cell marker. This antibody reacts with an antigen present in early thymocytes. The positive staining of this marker may represent a sign of early commitment to the T-cell lineage.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Beverley PC, et al. Eur J Immunol. 1981; 11:329-34
References 2:
Clevers H, et al. Eur J Immunol. 1988; 18:705-10
References 3:
Hedvat CV, et al. Hum Pathol. 2002; 33:968-74
References 4:
Karube K, et al. Am J Surg Pathol. 2003; 27:1366-74
CD1a is a non-polymorphic, major histocompatibility complex, class I-related cell surface glycoprotein (45 to 55 kDa) and is expressed in association with ?-microglobulin. In normal tissues, anti-CD1a reacts with cortical thymocytes, Langerhans cells, interdigitating cells, and rare antigen-presenting cells of the lymph node. CD1a positivity has also been seen in Langerhans cell histiocytosis (histiocytosis X), and a subset of pre-T lymphoblastic lymphoma/leukemia (cortical T LBL/L).
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP3622
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Krenacs L, et al. J Pathol. 1993; 171:99-104
References 2:
Angel CE, et al. Blood. 2009; 113:1257-67
References 3:
Emile JF, et al. Am J Surg Pathol. 1995; 19:636-41
References 4:
Stefano, AP et al. Br J Haematol. 1999; 105:394-401
Calponin is a 34 kD polypeptide that interacts with actin, tropomyosin, and calmodulin. It is involved in smooth muscle contraction mechanism and is restricted exclusively to smooth muscle tissue. Anticalponin has been found to be useful in staining myoepithelium and is, therefore, useful for differentiating benign sclerosing adenosis of the breast from infiltrating ductal carcinoma. Calponin positivity has also been noted in malignant myoepithelioma and pleomorphic adenoma3 of salivary gland origin, as well as angiomatoid malignant fibrous histiocytoma.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
EP798Y
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Wang NP, et al. Appl Immunohistochem. 1997; 5:141-151
References 2:
Nagao T, et al. Cancer. 1998; 83:1292-9
References 3:
Savara AT, et al. Mod Pathol. 1997; 10:1093-1100
References 4:
Fanburg-Smith JC, et al. Hum Pathol. 1999; 30:1336-43
References 5:
Hornick JL, et al. Am J Surg Pathol. 2003; 27: 1183-96
Calponin is a 34-kD actin filament-associated regulatory protein that interacts with actin, tropomyosin, and calmodulin. It is involved in the smooth muscle contraction mechanism and is restricted exclusively to smooth muscle tissue and myoepithelial cells. Anti-calponin has been found to be useful marker for differentiating benign sclerosing lesions of the breast from invasive carcinoma.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
CALP
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Anti-caldesmon is a regulatory protein found in smooth muscle and other tissues which interacts with actin, myosin, tropomyosin, and calmodulin. Anti-caldesmon antibody labels smooth muscle and tumors of smooth muscle, myofibroblastic, and myoepithelial differentiation. Anti-caldesmon has also been used to differentiate epithelioid mesothelioma from serous papillary carcinoma of the ovary.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
E89
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Miettinen M, et al. Arch Pathol Lab Med. 2006; 130:1466-78
References 2:
Watanabe K, et al. Hum Pathol. 1999; 30:392-6
References 3:
McCluggage WC. Adv Anat Pathol. 2004; 11:162-71
References 4:
Comin CE, et al. Am J Surg Pathol. 2006; 30:463-9
References 5:
Comin CE, et al. Am J Surg Pathol. 2007; 31:1139-48
Immunohistochemical staining with anti-calcitonin antibody has proven to be an effective way of demonstrating calcitonin-producing cells in the thyroid. C-cell hyperplasia and medullary thyroid carcinomas stain positive for calcitonin. Studies of calcitonin have resulted in the identification of a wide spectrum of C-cell proliferative abnormalities.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Matias-Guiu X, et al. Endocr Pathol. 2014; 25:21-9
References 2:
Fisher S, et al. Arch Pathol Lab Med. 2008;132:359-72
Carbohydrate Antigen 19-9 (CA19-9) is a sialylated Lewis A blood group antigen. It is synthesized by glycosyltransferases and has been identified as a component of gangliosides, glycoproteins and mucins. Anti-CA19-9 reacts with epithelial cells of normal pancreas, stomach, and colon as well as various adenocarcinomas, including pancreatic, gastric, and colorectal adenocarcinomas.
Antibody Isotype:
IgM
Monosan Range:
MONOSAN Ready To Use
Clone:
121SLE
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Encabo G, et al., Bull cancer (Paris) 1986;73:256-9
References 2:
Wu E, et al. Clin Adv Hematol Oncol. 2013; 11:535
References 3:
Partyka K, et al. Proteomics. 2012; 12:2212-20
References 4:
Remmers N, et al. Clin Cancer Res. 2013; 19:1981-93
The antibody reacts with epithelioid malignancies of the ovary, papillary serous carcinoma of the cervix, adenocarcinoma of the endometrium, clear cell adenocarcinoma of the bladder, and epithelioid mesothelioma. The antigen is formalin resistant, permitting the detection of ovarian cancer by immunohistochemistry, although serum assays for this protein are widely used to monitor ovarian cancer. MON 3211 also reacts with antigens in seminal vesicle carcinoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
OC125
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Kabawat S, et al. Int J Gynecol Pathol. 1983; 2:275-285
References 2:
Davis H, et al. Cancer Res. 1986; 46:6143-6148
References 3:
Zhou C, et al. Am J Surg Pathol. 1998; 22:113- 20
References 4:
Mylonas I, et al. Anticancer Res. 2003; 23:1075-80
References 5:
Fukazawa I, et al. Arch Gynecol Obstet. 1988; 243:41-50
Beta-catenin is a 92 kD protein normally found in the cytoplasm of the cell in the submembranous location. Mutations in the beta-catenin gene result in nuclear accumulation of this protein. Nuclear accumulation of this protein has been demonstrated in fibromatosis (desmoid tumors) of the breast and abdomen and, therefore, is useful in differentiating from other spindle cell neoplasms that may occur in these locations.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
14
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Alman BA, et al. Am J Pathol. 1997; 151:329-34
References 2:
Li C, et al. Am J Pathol. 1998; 153:709-14
References 3:
Abraham SC, et al. Hum Pathol. 2002; 33:39-46
References 4:
Montgomery E, et al. Am J Surg Pathol. 2002; 26:1296-301
BCL6 is a transcriptional regulator gene which codes for a 706-amino-acid nuclear zinc finger protein. In normal tissue these antibodies have strong nuclear staining for a subset of B-lymphocytes, mostly located in germinal centers (GC). BCL6 antibodies stain malignant cells in follicular lymphoma, diffuse large B-cell lymphomas, Burkitt lymphoma,4 classical Hodgkin lymphoma, as well as majority of tumor cells in nodular lymphocyte predominant Hodgkin lymphoma. BCL6 expression has been also seen in anaplastic large cell lymphomas (ALCL)
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
GI191E/A8
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
BCL2 is a protein associated with apoptosis regulation produced by the bcl-2 gene, located on chromosome 14q32.BCL2 is comprised of an alpha (239 amino acids) and beta chain. BCL2 (and thus BCL2 alpha chain) is found in mitochondrial and nuclear membranes and in the cytosol rather than the cell surface. In normal lymphoid tissue, BCL2 antibody reacts with small B-lymphocytes in the mantle zone and many cells within the T-cell areas. Anti-BCL2 has shown consistent negative reaction on reactive germinal center B-cells and positive staining of neoplastic follicles in follicular lymphoma. This antibody is valuable when distinguishing between reactive and neoplastic follicular proliferation in lymph node biopsies. This antibody may also be used in distinguishing between those follicular lymphomas that express BCL2 protein and the small number in which the neoplastic cells are BCL2 negative.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN Ready To Use
Clone:
E17
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Cooper K, et al. Journal of Pathology. 1997; 182:307-10
References 2:
Chetty R, et al. J Clin Pathol. 1995; 48:1035-1038
Bcl-2 is the best characterized protein family involved in regulation of apoptotic cell death, consisting of anti-apoptotic and pro-apoptotic members. Bcl-2 is a useful marker for identifying neoplastic cells in follicular lymphoma. Antibodies specific for the Bcl-2 protein can be used to distinguish between reactive and neoplastic follicular proliferation in lymph node biopsies.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
124
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Tsujimoto, Y. Genes Cells. 1998; 3:697-707
References 2:
Gaulard P, et al. Am J Pathol. 1992; 140: 1089-95
References 3:
Wang T, et al. APMIS. 1995; 103:655-62
References 4:
West RB, et al. Am J Clin Pathol. 2002; 117:636-43
The antibody recognizes a human breast carcinoma associated glycoprotein BCA-225 (220-225kD). This protein differs in size and distribution from other breast carcinoma antigens. Anti-BCA-225 primary antibody labels breast cancer antigen 225 (BCA-225) in primary and metastatic breast carcinoma. BCA-225 was first identified in T47D breast carcinoma cells, but its expression in other carcinomas such as lung, kidney, ovary and endometrium has
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN Ready To Use
Clone:
Cu-18
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mesa-Tejada R, et al. Am J Pathol; 1988 130:305-14
Alpha-fetoprotein (AFP) is a fetal tumor-associated polypeptide of the albuminoid gene family that binds and transports molecules in addition to many other proposed functions. This secretory protein is synthesized primarily in the fetal liver whereas expression is repressed in adult liver.Anti-AFP has been immunohistochemically demonstrated in hepatocellular carcinoma (HCC) and shows no immunoreactivity in normal liver.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Mizejewski GJ et al. Exp Biol Med. 2001; 226:377-408
References 2:
Lazarevich NL et al.Biochemistry (Mosc). 2000; 65:117-33
References 3:
Yusof YA, et al. Anal Quant Cytol Histol. 2003; 25:332-8
Anaplastic lymphoma kinase (ALK) is a novel receptor protein-tyrosine kinase. ALK can create a fusion protein with a nuclear protein gene called nucleophosmin (NPM) via the amino terminus of NPM and the catalytic domain of ALK. The product of this fusion protein is oncogenic.1 Studies have found this chromosomal translocation in most anaplastic large-cell non-Hodgkin's lymphomas, making ALK a good marker for anaplastic large cell lymphomas
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
ALK-1
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
The antibody recognizes actin of skeletal, cardiac, and smooth muscle cells. It is not reactive with other mesenchymal cells except for myoepithelium. Muscle Specific Actin is a part of the actin family of proteins which are highly conserved, major components of the cytoskeleton. Anti-Muscle Specific Actin immunohistochemical reactivity is seen in skeletal, cardiac, and smooth muscle cells and can be seen in neoplasms with muscle differentiation such as leiomyomas and rhabdomyosarcomas. In contrast, antiMuscle Specific Actin reactivity is typically not seen in endothelial cells, connective tissues, carcinomas, melanomas, lymphomas and most nonmyogenic sarcomas
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
HHF35
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gown AM, et al. Am J Pathol. 1986; 125:191
References 2:
Schmidt RA, et al. Am J Pathol. 1988; 131:19-28
References 3:
Azumi N, et al.Mod Pathol. 1988; 1:469-74
References 4:
Rangdaeng S, et al. Am J Clin Pathol. 1991; 96:32-45
Smooth Muscle Actin is a part of the actin family of proteins which are highly conserved and form microfilaments. These filaments are one of the major components of the cytoskeleton. Anti-smooth muscle actin immunohistochemical reactivity is seen in smooth muscle cells, myofibroblasts and myoepithelial cells.
Antibody Isotype:
IgG1-k
Monosan Range:
MONOSAN Ready To Use
Clone:
1A4
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Cooke PH. A. J Cell Biol. 1976; 68:539-56
References 2:
Skalli O, et al. J Cell Biol. 1986; 103:2787-96
References 3:
Perez-Montiel MD, et al. Am J Dermatopathol. 2006; 28:105-11
ACTH or Adrenocorticotropic hormone is synthesized from pre-pro-opiomelanocortin (pre-POMC). ACTH is produced and secreted from corticotrophs in the anterior lobe (or adenohypophysis) of the pituitary gland. The anti-ACTH immunohistochemical reagent could be useful in the study of neoplastic and non-neoplastic pituitary diseases
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Pizarro CB, et al. Braz J Med Biol Res. 2004; 37:235-43
References 2:
Kageyama K, et al. Am J Med Sci. 2002; 324:326-30
References 3:
Fan X, et al. J Histochem Cytochem. 2002; 50:1509- 16
References 4:
Japon MA, et al. J Clin Endocrinol Metab. 2002; 87:1879-84
The immunohistochemical staining of Alpha-1-Antitrypsin is considered to be very useful in the study of inherited AAT deficiency, benign and malignant hepatic tumors and yolk sac carcinomas. Positive staining for A-1-Antitrypsin may also be used in detection of benign and malignant lesions of an histiocytic nature. Sensitivity and specificity of the results have made this antibody a useful tool in the screening of patients with cryptogenic cirrhosis or other forms of liver disease with portal fibrosis of uncertain etiology.
Monosan Range:
MONOSAN Ready To Use
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Callea F, et al. J Hepatol. 1986; 2:389-401
References 2:
Palmer PE, et al.Am J Clin Pathol. 1974; 62:350-4
References 3:
Palmer PE, et al. Cancer. 1980; 45:1424-31
References 4:
Raintoft I, et al. Hum Pathol. 1979; 10:419-24
References 5:
Ramsay AD, et al. Appl Immunohistochem Mol Morphol. 2008; 16:140-7
Uroplakins (UPs) are a family of transmembrane proteins (UPs Ia, Ib, II and III) that are specific differentiation products of urothelial cells. In non-neoplastic mammalian urothelium, UPs are expressed in the luminal surface plasmalemma of superficial (umbrella) cells, Uroplakin II/III cocktail is specific for tumors of urothelial origin and, when used in combination with other markers, can aid in the diagnosis of primary and metastatic tumors.
Uroplakins (UPs) are a family of transmembrane proteins (UPs Ia, Ib, II and III) that are specific differentiation products of urothelial cells. Uroplakins are markers of terminally differentiated urothelium. Uroplakin II (UPII) is a newly described sensitive marker for urothelial carcinoma (UC). The expression profile of UPII in different types of UC and its utility in the diagnostic setting are needed.
Uroplakins (UPs) are a family of transmembrane proteins (UPs Ia, Ib, II and III) that are specific differentiation products of urothelial cells. In non-neoplastic mammalian urothelium, UPs are expressed in the luminal surface plasmalemma of superficial (umbrella) cells, forming complexes of 16nm crystalline particles. UPIII is specific for tumors of urothelial origin and, when used in combination with other markers, can aid in the diagnosis of primary and metastatic tumors.
Isocitrate dehydrogenase 1 (IDH1) is a 46 kDa NADPdependent enzyme, which catalyzes the decarboxylation of isocitrate into ?-ketoglutarate. IDH1 may also play a role in the prevention of oxidative damage, and the shuttling of proteins to peroxisomes. It is widely reported that mutations in IDH1 result in multiple forms of gliomas. The Arginine to Serine substitution at amino acid 132 is seen in gliomas, abolishes magnesium binding and alters enzyme activity so that isocitrate is no longer converted to alpha-ketoglutarate but instead alpha-ketoglutarate is converted to R(-)-2-hydroxyglutarate.
MAPKs are involved in directing cellular responses to a diverse array of potentially harmful stimuli, such as mitogens, osmotic stress, heat shock, proinflammatory cytokines, but also growth factors (mammals). Possibly located exclusively in the cell nucleus, they regulate cell functions including proliferation, gene expression, differentiation, mitosis, cell survival, and apoptosis. In order to become active, they require usually multiple phosphorylation events in their activation loops, including phosphorylation by MAP2 kinases (Ste-7 kinases), which in turn are phosphorylated by the MAP3 kinase family, of which many are located at the cell membrane. Thus through this pathway, stimuli can effectively be conveyed from the cell membrane to the nucleus. Inactivation of MAPKs takes place by several phosphorylases, including dedicated phophorylases.
EBS-T-059 recognizes an oncofetal antigen of 220kDa, identified as a tumor-associated glycoprotein (TAG72) with properties of a mucin and usually expressed by adenocarcinomas, but not by mesotheliomas and thus helps in the differential diagnosis of malignancies of the lung. The combined use of anti-TAG-72 and anti-GCDFP-15 is valuable in the diagnosis of apocrine carcinoma. TAG72 is further used as serum marker for gastric cancer.
Bp53-12 reacts with an N-terminal epitope (aa 16-25) of both wild-type and mutated p53. This epitope is revealed in tissue sections only after formalin fixation. Mutation and/or allelic loss of p53 is one of the causes of a variety of mesenchymal and epithelial tumors. p53 Localizes in the nucleus, but is detectable at the plasma membrane during mitosis and when certain mutations modulate cytoplasmic/nuclear distribution.
Antibody Isotype:
IgG2a-A
Monosan Range:
MONOSAN
Clone:
Bp53-12
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bártek J. et al, J Pathol. 169(1):27-34 (1993)
References 2:
Vogelstein and Kinzler, Cell 70: 523-526, (1992)
References 3:
Hollstein et al, Science 253: 49-53: (1991)
References 4:
Lane, D.P, Nature 358: 15-16: (1992)
References 5:
Donehower et al, Biochemic. Biophys. Acta 1155: 181-182, (1993)
POH-1 detects the three bands within the 34kDa region corresponding to the p34 protein cyclin dependent kinase 1 (cdk1) and its cleavage products. It is positive in immunoblotting on HeLa cell lysate and negative on fibroblast (LEP strain) cell lysate. A cdc2 homolog, the cdk2 protein kinase, does not react with POH-1. Activated cdk1 / p34cdc2 performs specific functions during mitosis, including nuclear envelope breakdown and chromosome condensation.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
POH-1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Luká, J, et al. Eur. J. Biochem., 207: 169-176 (1992)
47-8D3 reacts with macrophages and detects the well-known leukocyte L1, cystic fibrosis antigen. Detecting a single protein band of 14 kDa in Western blots of lysates of human monocytes and granulocytes, the antigen was identified as the calcium-binding protein MRP14, which is a member of the S100 family involved a.o. in regulating the cell cycle. MRP14 is also implicated in the abnormal differentiation of myeloid cells in the stroma of cancer. It is further found on squamous mucosal epithelia. When associated with MRP8 it forms the heterodimer calprotectin.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
47-8D3
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Flavell DJ. et al., J. Histochem. Cytochem. 35: 1217-1226 (1987)
References 2:
Facchetti F. et al., Am. J. Clin. Pathol. 92: 42-50 (1989)
References 3:
Bardadin KA. et al., J. Pathol. 164: 253-259 (1991)
References 4:
Goebeler M. et al., J. Leukocyte Biol. 55: 259-261 (1994)
EBS-T-007 reacts with paracaspase MALT1, crucial for B- and T-cell activation/proliferation and activation of transcription factor NF-?B. It has an N-terminal death domain, two central immunoglobulin-like domains involved in the binding to the B-cell lymphoma 10 (BCL10) protein and a caspase-like domain. MALT1 and BCL10, can both be found translocated in MALT lymphoma. In such cases either BCL10 or MALT1 or both are highly expressed, depending on the site of translocation. Normal cells and MALT lymphomas lacking translocations exhibit much lower levels of expression.
EBS-T-006 reacts with MADER or Melanoma-Associated Delayed Early Response protein, encoded by the NAB2 gene, belonging to the family of NGFI-A binding proteins. They modulate transcription induced by some members of the EGR (early growth response) family of transactivators. NAB proteins can form homo- or hetero-multimers with other EGR or NAB proteins through a conserved N-terminal domain, and repress transcription through two partially redundant C-terminal domains.
EBS-T-005 reacts with MADER or Melanoma-Associated Delayed Early Response protein, encoded by the NAB2 gene, belonging to the family of NGFI-A binding proteins. They modulate transcription induced by some members of the EGR (early growth response) family of transactivators. NAB proteins can form homo- or hetero-multimers with other EGR or NAB proteins through a conserved N-terminal domain, and repress transcription through two partially redundant C-terminal domains.
EBS-C-003 recognizes Ku protein or XRCC5/6 (X-ray repair cross complementing protein 5/6), involved in Pol II-directed transcription by virtue of its DNA binding activity, serving as the regulatory component of the DNA-associated protein kinase that phosphorylates Pol II and transcription factor Sp. Ku proteins also activate transcription from the U1 small nuclear RNA and the human transferrin receptor gene promoters. It serves as autoantigen in patients with rheumatic diseases.
MFG-06 reacts with a glycoprotein of 40-45 kDa, known as Milk fat globule-EGF factor 8 protein (MFG-E8), lactadherin, p47 or milk fat globule 1 antigen. It is present on normal epithelial cells in various organs and considered a differentiation marker in carcinomas. It contains one EGF-like domain and 2 F5/8 type C domains. Functioning as a specific ligand for Integrin ?5 and Integrin ?3, MFG-E8 is thought to be involved in gamete interactions and cell attachment, possibly playing a role in fertilization and apoptosis. Additionally, MFG-E8 binds to rotavirus and inhibits its replication, thereby protecting the cell from viral infection. Overexpression of MFG-E8 is associated with breast cancer, suggesting that MFG-E8 may be related to tumorigenesis or progression. Among testicular carcinomas, seminomas are negative.
IPO-38 reacts with a 12-14 kDa protein, as found in Western blots of Raji cells, and appears in the mitotic cycle earlier than Ki-67. Lymphocytes, induced to early G1 phase by 12h exposure to PHA, will become positive while non-stimulated lymphocytes remain negative. Mononuclear cells and granulocytes of healthy donors are negative, while various forms of leukemia and lymphoma including Hodgkins disease are positive for IPO-38, as are many solid tumors such as some breast, gastric and colonic cancers for which it may serve as tumor progression marker.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
IPO-38
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Sidorenko, S.P. et al., Gematol. Transfuziol. 35: 19-22 (1990)
References 2:
Sidorenko, S.P. et al., Experem. Oncol. 16: 145-150 (1994)
References 3:
Thosaporn W., et al., Oral Dis. 10(1): 22-6 (2004)
References 4:
Hao Y. et al.,J Proteome Res. Sep;7(9): 3668-77 (2008)
References 5:
Makohon N.V. et al., Fiziol Zh. 54(6): 49-57 (2008)
7B6 reacts with CRASH, a 308 AA glycoprotein sharing 32% homology with human asparaginase and identical to Asparaginase-Like Protein (ALP) found in rat sperm. CRASH is found in a variety of tumors but only occasionally in normal tissues. Especially ovarian cancers are highly positive. CRASH is further found in brain tumors, some colonic cancers, some lung cancers, some prostate cancers, uterine cancers, some breast cancers and some thyroid cancers. It was not found in serum. In Western blot 3 bands are demonstrated of 45 kDa, 28 kDa and 15 kDa.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
7B6
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Weidle, UH et al, Anticancer Res. 29(4): 951-63 (2009).
4D11 reacts with CRASH, a 308 AA glycoprotein, sharing 32% homology with human asparaginase and identical to Asparaginase-Like Protein (ALP) found in rat sperm. CRASH is found in a variety of tumors but only occasionally in normal tissues. Especially ovarian cancers are highly positive. CRASH is further found in brain tumors, some colonic cancers, some lung cancers, some prostate cancers, uterine cancers, some breast cancers and some thyroid cancers. It was not found in serum. In Western blot 3 bands are demonstrated of 45 kDa, 28 kDa and 15 kDa.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
4D11
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Weidle, UH et al, Anticancer Res. 29(4): 951-63 (2009).
121SLE reacts with CA19-9 (>400 kDa) or sialyl Lea structure, which is synthesized from type 1 blood group precursor chains and is present in individuals expressing the Lea and/or Leb blood group antigens. 121SLE also binds to some extend to the afuco version of SLe a (LSTa; CA50). In normal tissues, CA19-9 is present in ductal epithelium of the breast, kidney, salivary, gland and sweat glands. Its expression is greatly enhanced in serum as well as in the majority of tumor cells in gastrointestinal (GI) carcinomas, including adenocarcinomas of the stomach, intestine and pancreas. 121SLE was typed in the ISOBM TD-6 workshop.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
121SLE
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Rye PD. et al, Tumor Biol 19 (5): 390-420, (1998).
References 2:
Christopher MG. et al, Cytojournal 8: 7 (2011)
References 3:
Rouger et al. Blood transfusion and immunohematol. 30(5): 353-720 (1987)
EBP-333 reacts with C/EBP? or CCAAAT/enhancer-binding protein beta, a transcription factor which is not only critical for normal macrophage functioning and differentiation, but affects a variety of other factors as well, such as cytokines (IL-6; IL-4; IL-5 and TNF-alpha), neurotransmitters and other neuronal factors and processes like muscle repair and the development of multi drug resistance in tumors (P-glycoprotein). Observations have implied that manipulation of the transcription factors involved may make it possible to modulate multidrug resistance, while leaving normal function of P-glycoprotein intact.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBP-333
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Couturier, C et al, Arterioscler Thromb Vasc Biol. 20(12): 2559-65 (2000)
MoBU-1 is reactive with cells containing incorporated 5-bromodeoxyuridine (BrdU) showing a clear, nucleus confined, speckled pattern. BrdU is an analogue of thymidine and can be introduced to proliferating cells in vitro or in vivo, which in turn incorporate BrdU into the DNA during S phase, prior to cell division. Immunocytochemical staining with MoBU-1 will show the degree of proliferation, detecting nucleated cells which have incorporated BrdU in place of thymidine into their DNA.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
MoBU-1 (85-2C8)
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Harms et al. Acta Histochemica, Suppl. Band 36, 353-359 (1988)
Storage of tissue samples. The aluminium cryo vials with screw caps can be used for storage of tissue samples. The cryo tubes are suitable for storage of tissue samples in liquid nitrogen. Suitable for deepfreezing till -196°C. Autoclavable at 121°C . Available in 3ml (PA6003) and 15ml (PA6015) (sets of 100 pieces).
Storage of cryo tubes in liquid nitrogen: 1) Place the tissue in the cryo tube, 2) Close the cryotube tighly and place the cryotube in the liquid nitrogen for 1 minute, 3) The cryo vials can be stored at -80°C. Use of cryo tube in combination with isopentane: 1)Place a cup with isopentane in the liquid nitrogen; after approximately 2 minutes the isopentane clot and have a temperature of -160°C. 2)Place the tissue sample or biopsy in the isopentane for 30 seconds, 3) Place the tissue in the cryo vials and store at -80°C.
Reagent preparation:
Make sure that the cap and the vial have the same temperature because the material could slightly shrink or expand under influence of excessive temperature differences.
Expected results:
Long term storage of tissue
Precautions:
Wear a face mask at all times, Check if the screw treads and the lid are okay, Screw the lid on tightly. Attention: AVOID EXCESSIVE TEMPERATURE CHANGES, the cap could explode from the cryo tube.
Storage of tissue samples. The aluminium cryo vials with screw caps can be used for storage of tissue samples. The cryo tubes are suitable for storage of tissue samples in liquid nitrogen. Suitable for deepfreezing till -196°C. Autoclavable at 121°C . Available in 3ml (PA6003) and 15ml (PA6015) (sets of 100 pieces).
Storage of cryo tubes in liquid nitrogen: 1) Place the tissue in the cryo tube, 2) Close the cryotube tighly and place the cryotube in the liquid nitrogen for 1 minute, 3) The cryo vials can be stored at -80°C. Use of cryo tube in combination with isopentane: 1)Place a cup with isopentane in the liquid nitrogen; after approximately 2 minutes the isopentane clot and have a temperature of -160°C. 2)Place the tissue sample or biopsy in the isopentane for 30 seconds, 3) Place the tissue in the cryo vials and store at -80°C.
Reagent preparation:
Make sure that the cap and the vial have the same temperature because the material could slightly shrink or expand under influence of excessive temperature differences.
Expected results:
Long term storage of tissue
Precautions:
Wear a face mask at all times, Check if the screw treads and the lid are okay, Screw the lid on tightly. Attention: AVOID EXCESSIVE TEMPERATURE CHANGES, the cap could explode from the cryo tube.
p63 is a type II integral membrane protein predominantly localized in the rough endoplasmic reticulum. p63 is reported to be expressed in a number of normal tissues including proliferating cells of the epithelium, cervix, urothelium and prostate. p63 is also reported to be expressed in most poorly differentiated squamous cell carcinomas.
Antibody Isotype:
IgG1, kappa
Monosan Range:
MONXtra
Clone:
7JUL
Concentration:
Greater than or equal to 208 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Mahalingam M et al. Modern Pathology. 2010; 23:713-719
References 2:
Shah VI et al. Histopathology. 2006; 48:683-691
References 3:
Yen CC et al. World Journal of Gastroenterology. 2005; 11(9):1267-1272
References 4:
Bilal H et al. The Journal of Histochemistry and Cytochemistry. 2003; 51(2):133-139
This monoclonal antibody recognizes both wild type and mutant forms of human p53 protein under denaturing and non-denaturing conditions. The epitope recognized by clone DO-7 can be destroyed by prolonged fixation in buffered formalin. The heat induced epitope retrieval technique may improve staining in some cases.
Antibody Isotype:
IgG2b
Monosan Range:
MONXtra
Clone:
DO-7
Concentration:
Greater than or equal to 22 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Tiniakos DG et al. Cytopathology. 1996; 7(3): 178186
References 2:
Yoshida T et al. Journal of Pathology. 2003; 199(2):166175
References 3:
Burns ASYW et al. British Journal of Cancer. 2002; 86(7):11171123
References 4:
Tweddle DA et al. American Journal of Pathology. 2001; 158(6): 20672077
References 5:
Fernando SS et al. International Journal of Surgical Pathology. 2000; 8(3):213222
Epidermal growth factor receptor (EGFR) is a transmembrane protein receptor of 170 kD with tyrosine kinase activity. Increased levels of EGFR are reported to be linked with malignant transformation of squamous cells eg in squamous cell carcinoma of the lung, head, neck, skin, cervix and esophagus. EGFR may also play a role in the development and progression of hepatocellular carcinomas where recurrence rates are higher in EGFR-positive cases. This correlation has similarly been reported in colorectal cancers where EGFR, produced by tumor cells, plays an important role in the invasiveness and proliferation of colorectal cancers. The majority of published studies of EGFR expression in human breast cancer has similarly shown an association with EGFR expression where it is inversely related to estrogen receptor status.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
EGFR.113
Concentration:
Greater than or equal to 26 mg/L
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Lodge AJ et al. Journal of Clinical Pathology. 2003; 56(4):300304
References 2:
Sriplakich S et al. BJU Int. 1999; 83(4):498503
References 3:
Inoue K et al. Acta Med Okayama 1998; 52(6):305310
References 4:
Tungekar MF and Linehan J. Journal of Clinical Pathology. 1998; 51:583587
Thyroglobulin is a heavily glycosylated protein of 670kD composed of two identical subunits and is synthesized by the follicular epithelial cells of the thyroid. Thyroglobulin provides iodination sites for the formation of the thyroid hormones.
Antibody Isotype:
IgG2a
Monosan Range:
MONXtra
Clone:
1D4
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Male DK et al. Immunology. 54: 419427 (1985)
References 2:
Shepherd PS et al. European Journal of Nuclear Medicine. 10: 291295 (1985)
References 3:
Chan CTJ et al. Clinical and Experimental Immunology. 70: 516523 (1987)
Mouse anti-Progesterone Receptor (A/B Forms), clone16 and SAN27
Antibody Type:
Monoclonal
Host Animal:
Mouse
Species Reactivity:
human
Immunogen:
Prokaryotic recombinant protein corresponding to the N-terminal region of the A form of the human progesterone receptor generating clone 16 and a prokaryotic recombinant protein corresponding to the 164 amino acid N-terminal region unique to the B form of the progesterone receptor generating clone SAN27.
The human progesterone receptor (PR) is expressed as two isoforms, PRA (94 kD) and PRB (114 kD), which function as ligand-activated transcription factors. In vitro studies have indicated that PRA and PRB can activate different target genes and that PRA, in some circumstances, may act as a dominant inhibitor of the function of PRB and other steroid hormone receptors. PRA and PRB are both expressed in normal breast. Most endometrial carcinomas, however, are reported to express only one isoform with either PRA or PRB being expressed. The cocktail has been formulated using two clones, clone 16, specific for PRA, and SAN27, specific for PRB.
The human progesterone receptor (PR) is expressed as two isoforms, PRA (94 kD) and PRB (114 kD), which function as ligand-activated transcription factors. These two isoforms are transcribed from distinct estrogen receptor (ER)-inducible promoters within a single copy PR gene. Clone 16 is specific for a region of the N-terminus of the A form of PR. The precise epitope has not been mapped but it reacts with both A and B forms of PR by Western blot but only with the A form by immunohistochemistry. This suggests that the epitope is inaccessible in the native folded B form of the protein.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
16
Concentration:
Greater than or equal to 324 mg/L
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Hungermann D et al. Journal of Pathology 2002; 198: 487494
Estrogen receptor (ER) content of breast cancer tissue is an important parameter in the prediction of prognosis and response to endocrine therapy. The introduction of highly specific monoclonal antibodies to ER has allowed the determination of receptor status of breast tumors to be carried out in routine histopathology laboratories.
Antibody Isotype:
IgG1
Monosan Range:
MONXtra
Clone:
6F11
Concentration:
n/a
Storage buffer:
Tissue culture supernatant with Sodium azide
Storage:
2-8°C
References 1:
Bevitt DJ et al. Journal of Pathology 1997; 183(2), 228232
References 2:
Kaplan, PA et al. Am J Clin Pathol 2005: 276280
References 3:
Zafrani B et al. Histopathology 2000; 37(6), 536545
References 4:
Harvey JM et al. Journal of Clinical Oncology 1999; 17(5), 14741481
References 5:
Khan SA et al.European Journal of Cancer 2000; 36(Suppl 4), S27S28
p40 is a relatively unknown antibody that recognizes ?Np63-a p63 isoform suggested to be highly specific for squamous/basal cells. In a recent study, p40 is equivalent to p63 in sensitivity for squamous cell carcinoma, but it is markedly superior to p63 in specificity1, which eliminates a potential pitfall of misinterpreting a p63-positive adenocarcinoma or unsuspected lymphoma as squamous cell carcinoma. These findings strongly support the routine use of p40 in place of p63 for the diagnosis of pulmonary squamous cell carcinoma. Postive control Prostate
BLA36 is a developmentally regulated 36 kDa antigen expressed on the plasma membrane of B lymphocytes, Reed-Sternberg, and mononuclear Hodgkins cells. It also gives strong staining of B cell lymphomas including follicular center cell lymphomas (large and small cell types), mantle zone lymphomas, and immunoblastic lymphomas.
EBS-T-001 reacts with BCL10. Having an N-terminal caspase recruitment domain (CARD), BCL10 can induce apoptosis and activate NF-?B. It is found on subpopulations of normal B and T cells, and is associated with MALT1, a paracaspase that, like BCL10, can be found translocated in MALT lymphoma. In such cases either BCL10 or MALT1 or both are highly expressed, depending on the site of translocation. MALT lymphomas lacking this translocation exhibit much lower levels of expression. BCL10 has been shown to be functionally conserved all the way back to zebrafish
Monoclonal antibody HPV-4G3 reacts with HPV-18 E6 peptide. Infection with specific types of HPV has been associated with an increased risk of developing cervical neoplasia. HPV types 6 and 11 have been associated with relatively benign diseases such as genital warts but types 16 and 18 are strongly associated with cervical, vaginal, and vulvar malignancies.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
HPV-4G3
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Beldermann, F. Virusprotein E6 des humanen Papillomavirus HPV-16, Thesis, University of Heidelberg, Germany (1995).
Monoclonal antibody HPV-4B12 reacts with HPV-16 E6 peptide (MHQKRTAMFQDPPQERPRKLPQLC). Infection with specific types of HPV has been associated with an increased risk of developing cervical neoplasia. HPV types 6 and 11 have been associated with relatively benign diseases such as genital warts but types 16 and 18 are strongly associated with cervical, vaginal, and vulvar malignancies.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
HPV-4B12
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Beldermann, F. Virusprotein E6 des humanen Papillomavirus HPV-16, Thesis, University of Heidelberg, Germany (1995).
Monoclonal antibody HPV-13E2 reacts with HPV-16 E6 peptide (MHQKRTAMFQDPPQERPRKLPQLC). Infection with specific types of HPV has been associated with an increased risk of developing cervical neoplasia. HPV types 6 and 11 have been associated with relatively benign diseases such as genital warts but types 16 and 18 are strongly associated with cervical, vaginal, and vulvar malignancies
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
HPV-13E2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Beldermann, F. Virusprotein E6 des humanen Papillomavirus HPV-16, Thesis, University of Heidelberg, Germany (1995).
Membrane fusion is mediated by envelope glycoproteins for enveloped viruses like herpes simplex. Four of at least 10 viral glycoproteins are necessary and sufficient to facilitate fusion of herpes simplex to target cells. These four glycoproteins include glycoprotein B (gB), glycoprotein D (gD), glycoprotein H (gH) and glycoprotein L (gL). Fusion is dependent upon the expression of a gD receptor on target cell membranes.postive: HSV-1, Negative HSV-2.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-I-042
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bystricka, M, et al, Acta Virol. 43: 399-402 (1999)
Giardiasisis a diarrheal illness caused by a single celled microscopic protozoan parasite, Giardia lamblia, also known as Giardia intestinalis. Giardia lamblia exists in two forms, an active form called a trophozoite, and an inactive form called a cyst. The active trophozoite attaches to the lining of the small intestine and is responsible for causing the signs and symptoms of giardiasis. The trophozoite cannot live long outside of the body and spread of infection is via the cyst, which is excreted in the host's feces. When it is ingested, stomach acid activates the cyst, and the cyst develops into the disease causing trophozoite in the new host. Giardiasis is diagnosed by finding cysts or trophozoites in the feces.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-I-039
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Misra, V, et al, Indian J. Pathol. Microbiol. 49: 519-523 (2006)
EBS-I-025 reacts with a conserved repeat domain on LMP1 (AA 268-286), present in all EBV isolates. LMP1 is expressed in most viral latency stages in vitro and in vivo
EBS-I-024 reacts with a conserved domain on EBNA1 (AA 430-442), present in all EBV isolates. EBNA1 is expressed in all viral latency stages in vitro and in vivo.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-I-024
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Chen, MR et al. J. Virol. 67(8): 4875-85 (1993)
References 2:
Chen, MR et al. J. Biomed. Sci. 5(3): 173-9 (1998)
References 3:
Sivachandra, N, et al, J. Virol. 86(1): 60-68 (2012)
Ten to forty percent of the patients with acquired immunodeficiency syndrome (AIDS) develop cytomegalovirus (CMV) infections. In some patients with AIDS, CMV is detected in the bronchoalveolar lavage fluid (BALF), urine, and other specimens, even when there are no symptoms of CMV disease. An indicator of active CMV infection is needed to facilitate the diagnosis of CMV disease in patients with AIDS or HIV infection. CMV p65 antigen was detected in the leukocytes of both the peripheral blood and BALF during the early phase of CMV disease.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
CMV-227
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Hanabusa H, et al, Kansenshogaku Zasshi. 68(9): 1105-12 (1994)
Ten to forty percent of the patients with acquired immunodeficiency syndrome (AIDS) develop cytomegalovirus (CMV) infections. In some patients with AIDS, CMV is detected in the bronchoalveolar lavage fluid (BALF), urine, and other specimens, even when there are no symptoms of CMV disease. An indicator of active CMV infection is needed to facilitate the diagnosis of CMV disease in patients with AIDS or HIV infection. CMV p65 antigen was detected in the leukocytes of both the peripheral blood and BALF during the early phase of CMV disease
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
CMV-224
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Hanabusa H, et al, Kansenshogaku Zasshi. 68(9): 1105-12 (1994)
Ten to forty percent of the patients with acquired immunodeficiency syndrome (AIDS) develop cytomegalovirus (CMV) infections. In some patients with AIDS, CMV is detected in the bronchoalveolar lavage fluid (BALF), urine, and other specimens, even when there are no symptoms of CMV disease. An indicator of active CMV infection is needed to facilitate the diagnosis of CMV disease in patients with AIDS or HIV infection. CMV p65 antigen was detected in the leukocytes of both the peripheral blood and BALF during the early phase of CMV disease.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
CMV221
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Hanabusa H, et al, Kansenshogaku Zasshi. 68(9): 1105-12 (1994)
EBS-I-100 reacts with C. difficile Toxin A, but not with V. cholerae subunit a, V. cholerae toxin, Pseudomonas aeruginosa exotoxin A, H-LT, P-LT. C. difficile is a major nosocomial pathogen that causes antibiotic-associated colitis and mediates inflammatory diarrhea by releasing two large protein enterotoxins (toxin A and toxin B) that are able to disrupt intestinal epithelial cells via their transferase activity and ability to monoglucosylate members of the Rho family. C. difficile toxin A is a toxin that is composed of 39 repeats that are responsible for binding to intestinal epithelial cell surface carbohydrates. C. difficile toxin A causes significant apoptosis of colonocytes which contributes to the formation of ulcers and pseudo-membranes in a pathway that involves p38-dependent activation of p53 and induction of p21, leading to cytochrome c release and caspase-3 activation through Bak activation.
Antibody Isotype:
IgG3-K
Monosan Range:
MONOSAN
Clone:
EBS-I-100
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kim H, et al, Gastroenterology 129: 1875-1888 (2005)
References 2:
Carter JP, et al, Gut Microbes. 1(1): 5864 (2010)
EBS-I-002 reacts with a soluble excreted antigen in ELISA. This determinant is unaffected by frozen storage of specimens, unlike antibodies to flagellar antigens which require fresh cultured organism. Positive: C. jejuni, type 1 (K807, K858, K634) Type 2, C. coli, C. hyointestinalis, C lardis. Cross reacts with Staphylococcus aureus and weakly with Pseudomonas fluorescens. Negative: C. fetus, C. fetus intestinals, C. faecalis, H. pylori, Listeria monocytogenes, Actinomyces israelii, E. coli, Lactobacillus casei, Bacillus cereus, C. freundi, Salmonella Virchow, Streptococcus faecalis and Enterobacter aerogenes.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-I-002
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Baily E.L. et al. Mol. Ecol 24(1): 208-21 (2015)
References 2:
Haddock G et al. microbiology 156(10): 3079-84 (2010)
References 3:
Altekruse, SF, at al, Emerg Infect Dis. 5: 28-35 (1999)
EBS-I-001 reacts with a soluble excreted antigen in ELISA. This determinant is unaffected by frozen storage of specimens, unlike antibodies to flagellar antigens which require freshly cultured organisms.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-I-001
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Altekruse, SF, at al, Emerg Infect Dis. 5: 28-35 (1999)
EBS-I-014 reacts with flagellar core protein of Borrelia species including B.burgdorferi. B.burgdorferi is a species of bacteria of the spirochete class of the genus Borrelia causing Lyme disease, a vector-borne, multisystem inflammatory disease which is transmitted to humans by the bite of ticks of the Ixodes ricinus complex
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-I-014
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Busch, U. et al, J. Clin. Microbiol. 34: 1072-1078 (1996)
The CD74 cluster, established during the IVth and Vth Leukocyte Typing Workshops, comprises four species of proteins (MW 41/35/33 kDa), all coded by a single gene, consisting of nine exons. CD74 is expressed primarily by antigen presenting cells, such as B-lymphocytes (from before the pre-B cell stage to before the plasma cell stage), macrophages, and monocytes, and many epithelial cells. In tissue sections anti-CD74 show a binding pattern very similar to that of anti-HLA-DR. It binds to the peptide binding groove of newly synthesized MHC class II alpha/beta heterodimers and prevents their premature association with endogenous polypeptides. EBS-CD-041 epitope is localized in the extracellular domain of CD74.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-041
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Epstein A.L. et al. J. Immunol. 133: 1028-1036 (1984)
References 2:
Marder, R.J. et al. Lab. Invest. 52: 497-504 (1985)
References 3:
Lazova R. et al. Cancer 79: 2115-2124 (1997).
References 4:
Pich, A. et al. Eur J Basic Appl Histochem. 35(1): 81-9 (1991)
MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an ? and ? chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B-cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-O-111
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Thompson CJ et al., Human Immunol 6: 133-150 (1983)
CDw78 (also called Ba antigen, Leu21 or LO panB a) is present on some immature and some mature B-cells. The antigen appears on B-cell progenitors preceding CD10, CD19, CD22 and CD37. It is expressed on resting B-cells and reappears and persists in the cytoplasm and on the cell surface until cytoplasmic Ig appears. Its expression is greatly increased after B-cell activation in vitro. It is also found on tissue macrophages and on epithelial cells, but not on Tcells, NK-cells, monocytes, granulocytes, thymocytes or bone marrow stromal fibroblasts nor myeloid tissues.
Antibody Isotype:
IgG3-K
Monosan Range:
MONOSAN
Clone:
IPO-10
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Pinchouk V.G. et al, Anticancer Res. 8: 1377-1380 (1988)
CDw78 (also called Ba antigen, Leu21 or LO panB a) is present on some immature and some mature B-cells. The antigen appears on B-cell progenitors preceding CD10, CD19, CD22, and CD37. It is expressed on resting B-cells and reappears and persists in the cytoplasm and on the cell surface until cytoplasmic Ig appears. Its expression is greatly increased after B-cell activation in vitro. It is also found on tissue macrophages and on epithelial cells, but not on T-cells, NK cells, monocytes, granulocytes, thymocytes or bone marrow stromal fibroblasts nor myeloid tissues. 60-3G2 was typed at CD workshop IV.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
60-3G2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Pinchouk VG. et al, Anticancer Research 8: 1377-1380 (1988)
References 2:
Gluzman DF. et al, Tissue Antigens 33: 151 (1989)
References 3:
Sidorenko SP. et al, Neoplasma 39: 3-9 (1992)
References 4:
Moldenhauer et al, Leucocyte Typing IV, pp 155 162, (1989)
References 5:
Pezzuto et al, Leucocyte Typing IV, pp 165 174, (1989).
MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an ? and ? chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
Bra30
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Chorvath B et al. Neoplasma 34(4): 417-425 (1987)
References 2:
Horejsi V et al. Tissue Antigens 32(1): 6-11 (1988)
MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an ? and ? chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes.
Antibody Isotype:
IgG2-K
Monosan Range:
MONOSAN
Clone:
EBS-O-110
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Sparrow RL, et al., Transplantation 42: 647-652 (1986)
References 2:
Chorvath B et al. Neoplasma 34(4): 417-425 (1987)
References 3:
Horejsi V et al. Tissue Antigens 32(1): 6-11 (1988)
MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an ? and ? chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes.
Antibody Isotype:
IgG2b-K
Monosan Range:
MONOSAN
Clone:
LN-3
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Marder, R.L. et al. Lab. Invest. 52: 497-504 (1985)
References 2:
Andrade, R.E. et al. Human Pathology 19: 932-941 (1988)
References 3:
Azumi N. et al. Human Pathology 19: 1376-1382 (1988)
EBS-O-114 reacts with HLA Class II. The antibody was shown to strongly block cytotoxic activity of T4+ cytotoxic T cell clones. Distribution studies on cell lines suggested that EBS-O-114 is directed against a DQ rather than a DR antigen.
MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an ? and ? chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes.
Antibody Isotype:
IgG3-K
Monosan Range:
MONOSAN
Clone:
Bra14
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Chorvath B et al. Neoplasma 34(4):417-425 (1987)
References 2:
Horejsi V et al. Tissue Antigens 32(1):6-11 (1988)
MHC class II molecules are encoded by polymorphic MHC genes and consist of a non-covalent complex of an ? and ? chain. Helper T lymphocytes bind antigenic peptides presented by MHC class II molecules. MHC class II molecules bind 13-18 amino acid antigenic peptides. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM and -DO molecules regulate binding of exogenous peptides to class II molecules (HLA-DR) by sustaining a conformation that favors peptide exchange. The differential structural properties of MHC class I and class II molecules account for their respective roles in activating different populations of T lymphocytes.
Antibody Isotype:
IgG2b-K
Monosan Range:
MONOSAN
Clone:
BraFB6
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Babusikova O et al., Neoplasma 32(6): 657-62 (1985)
EBS-O-109 reacts with human ?-2 microglobulin, a 22 kDa protein, which associates noncovalently with the 44kDa ?1-chain of the HLA Class I complex found on all nucleated cells and on platelets. There is no reaction with erythrocytes, neither with non-human primate cells. The detection of ?-2 microglobulin in body fluids has been used as a tumor marker, renal failure marker and for monitoring patients with HIV infection.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-O-109
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Brodsky FM et al, lmmunol Rev 47: 3-61 (1979)
References 2:
Ryschich, E, et al, Clin Cancer Res. 11(2 Pt 1): 498-504 (2005)
Betts RL, et al. Monoclonal hybridoma antibodies: Techniques and applications. Edited by D. Hurrel. Uniscience series program. C.R.C. Press, Cleveland, OH: (1983), pp. 193-222
EP-4 recognizes the HLA-B27 cell surface antigen on human cells. HLA-B27 has been found to be highly associated (60%) with ankylosing spondylitis (Bekhterev's disease, MarieStrümpell disease or "bamboo spine"). While rheumatoid factor tests will be negative, testing for the presence of HLA-B27 in a patient can confirm the diagnosis of ankylosing spondylitis. Also postgonococcal arthritis and acute anterior uveitis are associated with HLA-B27.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EP-4
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Hansen JA et al. Hematol Oncol Clin North Am, 4(3): 507-515 (1990)
References 2:
El-Shabrawi, Y. et al. Ophthalmology 113: 695-700 (2006)
108-2C5 recognizes an intralocus determinant present on a limited number of HLA-A locusencoded gene products (HLA-A2, -A3, A28, -A29, -A30, -A31 and -Aw33). Furthermore, by testing its reactivity with HLA-A2 natural variants and mutants, the importance of amino acid residues 79 and/or 80 of the alpha1 domain was demonstrated in the formation of an intralocus HLA-A determinant.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
108-2C5
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Lozano, F. et al., Immunogenetics 30: 50-53 (1989)
References 2:
Lozano, F. et al., Tissue Antigens 35: 193-195 (1990)
References 3:
Domenech, N. et al., Human Immunol 30: 140-146 (1991)
References 4:
Ryschich, E, et al, Clin Cancer Res. 11(2 Pt 1): 498-504 (2005)
Bra23/9 reacts with a monomorphic determinant of human major histocompatibility (MHC) class I antigens (HLA-A, B and C). Human MHC class I antigens are expressed constitutively on all nucleated cells and platelets and are absent on erythrocytes. MHC class I antigens play a role in class I MHCassociated antigen presentation, inhibition of NK cell cytotoxicity, tumor surveillance, and tissue allotransplantation.
EBS-O-104 reacts with a monomorphic determinant of human major histocompatibility (MHC) class I antigens (HLA-A, B and C). Human MHC class I antigens are expressed constitutively on all nucleated cells lymphocytes such as lymphocytes, thymocytes, granulocytes, and bone marrow cells and also platelets but are absent on erythrocytes. MHC class I antigens play a role in class I MHCassociated antigen presentation, inhibition of NK cell cytotoxicity, tumor surveillance, and tissue allotransplantation.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-O-104
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Young NT et al. Hum Immunol 52(1): 1-11 (1997)
References 2:
Krensky AM et al, Transplant Proc 28(6): 3026-8 (1996)
References 3:
Hansen JA et al, Hematol Oncol Clin North Am 4(3): 507-515 (1990)
LN-6 reacts with vimentin, a 58kDa protein, and specifically with a non-hematopoietic epitope of vimentin. It shows no cross-reaction with other closely related intermediate filament proteins (IFP) such as desmin, keratin, neurofilament, and glial fibrillary acid protein. Vimentin is ubiquitously expressed in mesenchymal cells such as fibroblasts, smooth muscle cells, and endothelium. Antibody against vimentin is useful as part of an antibody panel for differential diagnosis of tumors of unknown origin. It does not react with leukocyte common antigen-positive tissues such as lymphomas and leukemias.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
LN-6
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Stathopoulos, E, et al, J. Histochem. Cytochem. 37: 1363-1370 (1989)
C11 reacts with keratins : 4,5,6,8,10,13 and 18. This is a broad-spectrum antibody, which has been reported to differentiate epithelial tumors from non-epithelial tumors. Many studies have shown the usefulness of keratins as markers in cancer research and tumor diagnosis.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
C-11
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Vojt?ek, B. et al. Folia Biol., 35(6): 373-382, (1989)
References 2:
Vojt?ek, B. et al. Neoplasma 37(3): 333-342 (1990)
References 3:
Kova?ík J. et al. Int. J. Cancer, Suppl. 3: 50-55, (1988)
References 4:
Kova?ík J. et al. J. Tumor Marker Oncol. 5: 219 (1990)
EBS-IF-007 is human specific and reacts with rod domain 2B, aa 311-335, of keratin 19 (40 kDa). Keratin 19 is expressed in sweat gland, mammary gland ductal and secretory cells, bile ducts, gastrointestinal tract, bladder urothelium, oral epithelia, esophagus, and ectocervical epithelium. Anti-keratin 19 reacts with a wide variety of corresponding epithelial malignancies.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-IF-007
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Böttger, V. et al., Eur. J. Biochem. 231: 475-485 (1995)
References 2:
Karsten et al., Eur.J.Cancer Clin. Oncol. 21(6), 733-740 (1985)
References 3:
Kasper et al., Histochemistry 89(4), 369-377 (1988)
C-04 Reacts with keratin 18 (45kDa) present in a wide variety of simple epithelia. It does not react with stratified squamous epithelia. It reacts with epithelial tumors of the gastrointestinal tract, lung, breast, pancreas, ovary, and thyroid.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
C-04
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bártek et al. J. of Pathol. 164: 215-24 (1991)
References 2:
Bártková et al. Neoplasma 38(4): 439-46 (1991)
References 3:
Lauerová et al. Hybridoma 7: 495-504 (1988)
References 4:
Taylor Papadimitriou et.al. J. Cell Sci. 94: 403-13 (1989)
References 5:
Kova?ík et al. J. Tumor Marker Oncol. 5: 219 (1990)
E3 reacts with keratin 17 (45 kDa). The antibody reveals myoepithelial cells, basal cells and proliferating epithelia in some benign epithelial tumors as well as malignant carcinomas.
EBS-IF-004 reacts specifically with keratin 14 (50kDa) in immunoblotting. In tissue sections EBS-IF-004 is positive with basal cells of non-keratinizing stratified epithelia, basal cells and suprabasal cells of the epidermis and gingiva, myoepithelial cells and squamous cell carcinomas.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-IF-004
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Ivanyi, D. et al. Am. J. Vet. Res. 53: 304-314 (1992)
References 2:
Ivanyi, D. et al. Cancer Res. 50(16): 5143-5152 (1990)
References 3:
Balm A.J.M. et al., Eur. Arch. Otorhinolaryngol. 253: 227-233 (1996)
EBS-IF-003 reacts with 53kDa (CK13) and 56.6kDa (CK10) cytokeratin proteins as indicated by immunoblotting. On formalin-fixed, paraffin-embedded tissue sections EBS-IF-003 recognizes only CK13. On cryostat sections EBS-IF-003 serves as differentiation-related marker of all stratified epithelia; it stains all suprabasal cells in both cornifying and noncornifying stratified epithelia and more differentiated cells of squamous carcinomas. In paraffin sections EBS-IF-003 does not stain CK10 positive, CK13 negative epithelia, as for example epidermis.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-IF-003
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Ivanyi D. et al, J. Pathol. 159: 7-12 (1989)
References 2:
Ivanyi D. et al, Am. J. Vet. Res. 53: 304-314 (1992)
References 3:
Ivanyi D. et al, Am. J. Vet. Res. 54: 1095-1102 (1993)
References 4:
Kozaki, M et al, J. Vet. Med. Sci. 63(1): 1-4 (2001)
C-51 reacts with keratin 8 (52.5 kDa) + 18 (45 kDa) polypeptides and recognizes all simple epithelia including glandular epithelium, for example thyroid, female breast, gastrointestinal tract, respiratory tract, and urogenital tract including transitional epithelium. All adenocarcinomas and most squamous carcinomas are positive but keratinizing squamous carcinomas are usually negative. This antibody is useful in demonstrating the presence of Paget cells; there is very little keratin 18 in the normal epidermis so only Paget cells are stained
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
C-51
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bártek et al. J. Pathol. 164: 215-24 (1991)
References 2:
Bártková et al. Neoplasma 38(4): 439-46 (1991)
References 3:
Wendl, J. et al. Anat. Histol Embryol. 164: 215-24 (1991)
References 4:
Thangappan, R et al. Cell. Polif. 42: 770-779 (2009)
C-46 reacts with keratins 7 (54kDa) and 17 (46kDa). On formalin-fixed paraffin embedded sections C46 reacts with keratin 7 only. Strongly positive on most simple epithelia except for stomach, small intestine and colon mucosa, hepatocytes, pancreatic acini, renal tubules, also positive on some non-cornifying epithelia.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
C-46
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bártek et al. J. of Pathol. 164: 215-24 (1991)
References 2:
Vojt?ek et al. Neoplasma 37(3): 333-342 (1990)
References 3:
Taylor Papadimitriou J. et. al. J. Cell Sci. 94: 403-13 (1989)
References 4:
Kova?ík et al. Int. J. Cancer Suppl. 3: 50-55, (1988)
References 5:
Kova?ík et al. J. Tumor Marker Oncol. 5: 219, (1990)
EBS-IF-001 Does not react with keratin polypeptides in immunoblotting. The specificity for keratin was established on the basis of antibody reactivity with intracytoplasmic intermediate filaments in epithelial, vimentin negative, human and feline cells. Distribution of the epitope strongly suggests that EBS-IF-001 reacts with keratins 5 and 14, most probably with a heterotypic complex formed by these keratins. EBS-IF-001 reacts positively with basal cells of all stratified epithelia, with myoepithelial cells and with most squamous cell carcinomas. It is useful in immunohistochemistry for typing of basal cells.
EBS-huLambda reacts with human immunoglobulin lambda light chain (22,5 kDa). EBShuLambda does not react with T cells, monocytes, granulocytes or erythrocytes.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-huLambda
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Baryshnikov A. et al. Gemat.Transf. (Russian) N8, 4-7 (1990)
References 2:
Martinova T.et al., In: Problems in med biotech. and immunol. Infect.diseases. 11, 182-186, (1996)
EBS-huKappa reacts with kappa light chain In mammals, the two light chains in an antibody are always identical, with only one type of light chain, kappa or lambda. The ratio of kappa to lambda is 70:30. However, with the occurrence of multiple myeloma or other B-cell malignancies this ratio is disturbed.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-huKappa
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Takahashi, H. et al. Pathol Res Prac 189, 300-311 (1993)
References 2:
Momose, H., et al. Hum. Pathol 23: 1115-1119 (1992)
SC-05 reacts with a reduction resistant epitope on 80 kDa human secretory component (both free and bound to SIgA). Secretory component is differentially expressed in epithelium, thus SC-05 can identify subpopulations of epithelial cells and epithelial differentiation. Secretory component negative cell lines are not stained with SC-05.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
SC-05
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kvale, D. et al, Int J Cancer 42(4): 638-641 (1988)
References 2:
Bartek, J. et al, Histochem 91(3): 235-244 (1989)
References 3:
Bartek, J. et al, Histochem J 22(10): 537-534 (1990)
EBS-huA recognizes the third constant domain (CH3) of the alpha chain of human IgA and reacts with both IgA1 and IgA2 isotypes and not with other types of immunoglobulins. IgA type antibodies are secreted by B-cells associated with mucosal epithelia and therefore indicate malignancy if found in lymphoid infiltrates at other locations. Detection of IgA antibodies to EBV in serum may indicate nasopharyngeal carcinoma. Rising IgA anti EBV levels are associated with progression of disease.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-huA
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Biewenga J. et al, Mol. Immunol. 23: 761-767 (1986)
References 2:
Biewenga J. et al, Adv. Exp. Med. Biol. 216B: 1239-1249 (1987)
References 3:
Mestecky J. et al, J. Immunol. Meth. 193: 103-148 (1996)
References 4:
Oortwijn B.D. et al, Mol Immunol. 44(5):966-73 (2007)
VEGF-21 reacts with Vascular Endothelial Growth Factor, also known as Vascular Permeability Factor (VEGF/ VPF) and is the key mediator of angiogenesis. The MWs are 19- 22kDa (reducing) and 38kDa-44kDa (non-reducing). There are multiple isoforms of VEGF containing 206-, 189-, 165-, and 121-amino acid residues. The smaller two isoforms, VEGF165 and VEGF121, are secreted proteins and act as diffusible agents, whereas the larger two remain cell associated. VEGF/VPF plays an important role in angiogenesis, which promotes tumor progression and metastasis. In addition to endothelial cells, VEGF and VEGF receptors are expressed on numerous non-endothelial cells including tumor cells.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
VEGF-21
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Tischer E. et al. J. Biol. Chem 266: 11947-11954 (1991)
UACA (Uveal Autoantigen with Coiled-coil domains and Ankyrin repeats) is a 1,416 amino acid nuclear membrane protein. It was originally identified as an autoantigen in patients with panuveitis, a characteristic of Vogt-Koyanagi-Harada disease, and in patients with Graves' disease. UACA was also later identified as Nucling, a mRNA differentially expressed in F9 embryonal carcinoma cells, and that is up-regulated during cardiac muscle differentiation. UACA appears to function as a pro-apoptotic protein that recruits the apaf-1- pro-caspase-9 complex for the induction of apoptosis to mediate the cell-death pathway.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
AE-5
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Yamada, K., et al. Biochem. Biophys. Res. Commun. 280: 1169-1176 (2001)
TU-02 reacts with the N-terminal domain of alpha-tubulin. Tubulin isotypes have been identified with tissue specific expression. Immunocytochemical studies using several mAbs revealed remarkable heterogeneity of tubulin within various nervous tissues. TU-02 reacts with tubulin in fixed tissues only, not in unfixed or live tissues or cells. Interphase microtubules are also stained by TU-02 in fixed tissues.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
TU-02
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Dráber, P. et. al. Eur.J.Cell.Biol. 41: 82-88 (1986)
References 2:
Dráber, P. et. al. Histochemistry 87: 151-155 (1987)
References 3:
Dráber, P. et. al. J. Cell Science 92: 519-528 (1989)
References 4:
Smertenko et al. Eur. J. Cell. Biol. 72: 104-112 (1997)
1E8-G6 reacts with TGF-alpha and shows no cross-reaction with EGF and the neuropeptide synenkephalin. 1E8-G6 is completely blocked by the peptide used for immunization. TGFalpha (aa50) is a growth factor with 33% homology to EGF, binds to EGFR, activates tyrosine phosphorylation of the receptor, and stimulates cell proliferation. It plays a role in tumor initiation by inducing the reversible transformed phenotype. In sections both cytoplasma and cell membranes are stained
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
1E8-G6
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bebók, Zs, et al. Breast Cancer Res.Treatm. 29: 229-235 (1994)
EBS-O-166 specifically reacts with tenascin-C, an extracellular matrix glycoprotein of 210 kD. It recognizes those forms of tenascin that are produced by both normal and hyperproliferative (also neoplastic) tissues. Tenascin/hexabrachion/cytotactin is an extracellular matrix glycoprotein, widely expressed during embryogenesis. In adults, it is restricted to certain epithelial-stromal interfaces and increases markedly in hyperproliferative diseases and in stroma of many neoplasms, including gliomas, breast, squamous and lung carcinomas.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-O-166
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Verstraeten, AA et al., Br. J. Derm. 127(6), 571-574 (1992)
PLAP is a tissue specific, throphoblast-derived, 58 kDa, glycosyl-phosphatidylinositol (GPI)- anchored, dimeric, Zn2+ metallated glycoprotein, only found in humans, orangutans and chimpanzees, that catalyzes the hydrolysis of phosphomonoesters into an inorganic phosphate and an alcohol. It is present in the placenta and serum of pregnant women and in high frequency in gynecological and testicular cancers and in lower frequency in other tumors. The three tissue-specific APs in humans, PLAP, germ cell AP (GCAP) and intestinal AP, are 90-98% homologous. Non tissue specific AP is found in kidney, liver and bone. H7E8 binds equally well to all common allelic variants (S,F, FS and I) of PLAP as to AP from normal human testis, while antibody F5C2 reacts with some samples of normal human testis only.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
H7E8
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Millan J.L. et al, Eur. J. Biochem. 136: 1-7 (1983)
PLAP is a tissue specific, throphoblast-derived, 58 kDa, glycosyl-phosphatidylinositol (GPI)- anchored, dimeric, Zn2+ metallated glycoprotein, only found in humans, orangutans and chimpanzees, that catalyzes the hydrolysis of phosphomonoesters into an inorganic phosphate and an alcohol. It is present in the placenta and serum of pregnant women and in high frequency in gynecological and testicular cancers and in lower frequency in other tumors. The three tissue-specific APs in humans, PLAP, germ cell AP (GCAP) and intestinal AP, are 90-98% homologous. Non tissue specific AP is found in kidney, liver and bone. F5C2 binds equally well to all common allelic variants (S,F, FS and I) of PLAP and to some variants of AP from normal human testis, while antibody H7E8 reacts with all variants of AP in normal human testis.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
F5C2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Millan J.L. et al, Eur. J. Biochem. 136: 1-7 (1983)
IPO-74 reacts with human leukemia cell line HL-60 and shows a band at MW 127 kDa (reduced) both in immunoprecipitation and Western blotting. In peripheral blood only subsets of B cells, T cells, neutrophilic and eosinophilic granulocytes are positive. All monocytes are positive. Cell lines U937 and K562 are positive, while Daudy, Raji, Jurkat and Molt-4 are negative, as are IL2-dependent T cell lines/clones.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
IPO-74
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Fleming MG, et al, J Cutan Pathol. 17(2): 77-81 (1990)
D11 reacts specifically with human monocytes and macrophages, in all sorts of tissues. The 125/135 kDa antigen is present on the cell membrane as well as within cytoplasmic structures including lysosomes, and is different from CD68. Among tumors, histiocytomas and histiocytic lymphomas are positive. In ALL, positive reaction with D11 indicates B-lineage derivation. AML are negative.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
D11
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Rudinskaya T.D. et al. Immunol Lett. 33(1): 1-7 (1992)
References 2:
Frenkel M.A. et al. Gematol Transfuziol. 40(4): 13-16 (1995)
References 3:
Tupitsyn N.N et al., Int J Cancer. 68(2): 160-163 (1996)
References 4:
Petrovichev N.N et al., Acta Cytol. 41(2): 357-363 (1997)
FR4A5 reacts with CD15 (220 kDa). CD15 contains the pentasaccharide lacto-n-fucopentaose III and plays a role in mediating phagocytosis, bactericidal activity, and chemotaxis. It is present on >95% of granulocytes including neutrophils and eosinophils and to a lesser degree on monocytes. In addition, CD15 is expressed in Reed-Sternberg cells and some epithelial cells. CD15 is occasionally expressed in large cell lymphomas of both B and T phenotypes which otherwise have a quite distinct histological appearance. It is also expressed on a wide variety of other tumor cells including myeloid leukemia, breast, colorectal, and lung cancer cells. Cross reactivity has been observed with Glc?1-6glc?1-4Glc, Tn, blood group H1, and maltose.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
FR4A5
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Manimala J.C. et al. Glycobiology. 17(8): 17C-23C (2007)
References 2:
Gildersleeve J. et al. Glycobiology. 18(0): 746 (2008)
Laminins are large hetero-trimeric, non-collagenous glycoproteins found in basement membranes and composed of ?, ?, and ? chains. A5 reacts specifically with ? chain 1. Alterations of basement membrane integrity, from local discontinuities up to complete loss, are described in many types of human and animal epithelial neoplasms.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
A5
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Ljubimov JY et. al. Cancer Res. 61(14): 5601-5610 (2001)
References 2:
Ljubimov AV et. al. Int J Cancer 50: 562-566 (1992)
The alpha interferons are involved in virus resistance in target cells for these viruses. They are known to block cell proliferation and to regulate MHC class I antigen expression. The IFN? family has over 20 genes and pseudogenes in two families (I and II), one with a mature length of 166aa and one of 172aa. Cells producing IFN? are lymphocytes, monocytes, macrophages and cell lines such as Namalwa and KGI. Bioassays for IFN? include cytopathic effect blocking, by viruses such as VSV, SFV and BMCV, on their target cells. A number of receptors for IFN? are now known and seem to be expressed on most cell types. 2-52 Is specific for human IFN?1 and does not cross react with human IFN?2.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
Feb-52
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kontsek, P. et al., Mol Immunol. 29: 863-870 (1992)
The alpha interferons are involved in virus resistance in target cells for these viruses. They are known to block cell proliferation and to regulate MHC class I antigen expression. The IFN? family has over 20 genes and pseudogenes in two families (I and II), one with a mature length of 166aa and one of 172aa. Cells producing IFN? are lymphocytes, monocytes, macrophages and cell lines such as Namalwa and KGI. Bioassays for IFN? include cytopathic effect blocking, by viruses such as VSV, SFV and BMCV, on their target cells. A number of receptors for IFN? are now known and seem to be expressed on most cell types. 2-48 Is specific for human interferon ? 1 and does not cross react with human interferon ? 2.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
Feb-48
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kontsek, P. et al. Mol Immunol. 29: 863-870 (1992)
EBS-O-148 can be used in ELISA, frozen sections, FACS and Western blot, however no method is known to date to retrieve the antigen in paraffin sections. Anti-hemoglobin antibodies can be used in diagnosis of anemias and as tumor marker in stool.
HE-182 reacts with human blood group H type 5 and type 6 as confirmed with Chembiomed synthetic oligosaccharide-BSA conjugate in ELISA. It is expressed on endothelial cells, epithelial cells and granulocytes. Increased expression of this antigen has been observed on some tumor tissues such as gastric carcinomas, urothelial carcinomas, and colon carcinomas
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
HE-182
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Rouher Ph et al. Blood transfusion and immuohaematology, Arnette France 30 (5): 353-720 (1987)
Until recently, immunological markers for myeloid cells have been lacking, especially those which identify different levels of cellular differentiation. The BM series provides a new panel of monoclonal antibodies which stain early precursor and mature forms of human myeloid cells. This panel of monoclonal antibodies reacts with antigenic determinants present in normal myeloid cells and leukemias of similar derivation. BM-2 recognizes a cytoplasmic antigen expressed in mature human granulocytes (polys) residing in lymphoid and non-lymphoid tissues. It does not react with any other cell type in human tissues.
618 Recognizes an epitope contained within the gelatin binding domain of human fibronectin Fibronectins, 220 kDa monomer; 440 kDa dimer, are present in basement membranes, interstitial connective tissue matrix, and blood. Cellular fibronectin is widely distributed in the stroma of many malignant tumors.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
618
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Ljubimov, A.V. et al. Exp. Cell Res. 165,53040 (1986)
616 Reacts with the N-terminal fibrin and heparin-binding domain. Specificity was established by Western blotting with purified 29 kDa domain from two different sources. Fibronectins, 220 kDa monomer; 440 kDa dimer, are present in basement membranes, interstitial connective tissue matrix, and blood. Cellular fibronectin is widely distributed in the stroma of many malignant tumors.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
616
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Grant, M B, et al. Diabetes, 47: 1311-7 (1998)
References 2:
Homandberg, G.A. et al. Osteoarthritis and Cartilage 6: 231244 (1998)
References 3:
Klueh, U, et al. Biomaterials 24(22) : 3877-84 (2003)
References 4:
Ljubimov, A.V. et al. Exp. Cell Res. 165: 53040 (1986)
References 5:
Mirmalek-Sani SH. et al. Biomaterials 165: 548895 (2013)
568 Reacts with the 8th type III repeat in the cell-binding domain of human fibronectin (220 kDa monomer; 440 kDa, dimer). Fibronectins are present in basement membranes, intestinal connective tissue matrix, and blood. Cellular fibronectin is widely distributed in the stroma of many malignant tumors.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
568
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Christensen, L. et al, APMIS 98(7): 615-623 (1990)
References 2:
Christensen, L. et al, APMIS suppl. 26: 1-39 (1992)
References 3:
Yong, JL. et al, Int J Clin Exp Pathol 3(2): 210-216 (2010).
TV-1 recognizes fibronectin in frozen and paraffin sections of human, pig, mouse and rat tissues. Specifically, it stains this extracellular adhesive glycoprotein in connective tissues and blood vessels. TV-1 does not recognize the soluble dimeric form of fibronectin (plasma fibronectin) but strongly stains matrix fibronectin in tissues. Cellular fibronectin is widely distributed in the stroma of many malignant tumors
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
TV-1 (2755-8, EP-5)
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Epstein, AL. et al. Cancer Res. 55(12): 2673-80 (1995)
HEB-29 shows specific staining of erythrocytes and vascular epithelium of blood group B and AB controls and no staining in group A or O controls. However, tumors in A or O individuals may carry B antigen and stain with HEB-29. HEB-29 is applicable for red cell agglutination, tissue staining and immunofluorescence tests.
HE-193 recognizes human blood group A (monofucosyl and difucosyl A antigens with chain types 1, 2, 3, 4, 5, 6,) and Forssmann antigen, which is normally not found in humans, but can appear on malignancies. De novo or increased expression of these antigens have been observed on some tumor tissues such as gastric carcinomas, urothelial carcinomas, and colon carcinomas. HE-193 does not react with normal tissue sections of donors with blood group B and 0 but it reacts specifically with malignant tissues in these individuals. HE-193 is applicable for red cell agglutination, tissue staining and immunofluorescence tests
HE-10 preferably reacts with determinants of chain A type 3 and 4 and chain H type 3 and 4, but not with type 1 and 2 chain structures. It is not reactive with immunodominant A trisaccharide. Increased expression of this antigen has been observed on some tumor tissues such as gastric carcinomas, urothelial carcinomas, and colon carcinomas. HE-10 does not react with normal tissue sections of donors with blood group B and 0 but it reacts specifically with malignant tissues in these individuals. HE-10 is applicable for red cell agglutination, tissue staining and immunofluorescence tests.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
HE-10
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
N?mec M. et al. Vox Sang 52:125-8 (1987)
References 2:
Vanák J. et al. Neoplasma 36(4): 479-487 (1989)
References 3:
Tichý, M. et al. Neoplasma 37(4): 451-459 (1990)
References 4:
Tichý, M. et al. Acta Univ Palacki Olomuc Fac Med. 126: 57-69 (1990)
3-3A reacts with determinants of chain A type 3 and 4 and chain H type 3 and 4, but not with type 1 and 2 chain structures. It is not reactive with immunodominant A trisaccharide. Increased expression of this antigen has been observed on some tumor tissues such as gastric carcinomas, urothelial carcinomas, and colon carcinomas. 3-3A does not react with normal tissue sections of donors with blood group B and 0 but it reacts specifically with malignant tissues in these individuals.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
3-3A
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al. Biochem. J. 254: 185-193 (1988)
References 2:
Yasumsds, I. et al. Glycoconjugate J. 3: 187-202 (2000)
References 3:
Rouger et al. Blood transfusion and immunohematol. 30(5): 252-720 (1987)
A-493 react with adiponectin, an adipocytokine. Adipocytokines are hormones produced in adipose tissue. Adiponectin is abundantly present in plasma and has insulin like effect on glucose levels in the blood. Plasma adiponectin levels are low in insulin resistant patients who are obese, have diabetes mellitus type 2 or HIV-lipodystrophy. In women adiponectin levels tend to be higher than men, which may be due to androgens suppressing adiponectin levels. Furthermore adiponectin and leptin are both indicated in regulating body weight through direct action on the hypothalamus, influencing appetite. Obese people have low adiponectin levels while levels in anorexia patients are high. Adiponectin acts as ligand for various receptors, two of which have been identified, one probably involved in carbohydrate assimilation, the other in tuning the rate of metabolism.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
A-493
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Dos Santos, E. et al, Oncol. Rep. 20: 971-977 (2008)
A-492 reacts with adiponectin, an adipocytokin; adipocytokines are hormones produced in adipose tissue. Adiponectin is abundantly present in plasma and has an insulin like effect on glucose levels in the blood. Plasma adiponectin levels are found in insulin resistant patients who are obese, have diabetes mellitus type 2 or HIV-lipodystrophy. In women adiponectin levels tend to be higher than in men, which may be due to androgens suppressing adiponectin levels. Furthermore adiponectin and leptin are both indicated in regulating body weight through direct action on the hypothalamus, influencing appetite. Obese people have low adiponectin levels while levels in anorexia patients are high. Adiponectin acts as ligand for various receptors, two of which have been identified, one probably involved in carbohydrate assimilation, the other in tuning the rate of metabolism.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
A-492
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Dos Santos, E. et al, Oncol. Rep. 20: 971-977 (2008)
This antibody is specific to a 45 kDa protein, which is identified as MyoD1. This antibody is specific to an epitope of amino acid 3-56 in the N-terminus of mouse MyoD1. This antibody does not react with myogenin, Myf5 or Myf6. MyoD1 stains the nuclei of myoblasts in developing muscle tissues. MyoD1 is not detected in normal adult tissue but is expressed strongly in the tumor cell nuclei of rhabdomyosarcomas.
The antibody reacts with mucin from small and large intestine and to a weaker extent with salivary and breast epithelia. Stomach, pancreas, kidney, and ovary epithelia are negative. Positive reaction of gastric cancer and colon cancer (antibody shows a relative high specificity for colon carcinoma). Reactivity on cultured cell lines: LS 174 T (colon cancer cell line). Positive control: Small intestine.
Mouse anti Human CD9 antibody, clone Bu16 detects human CD9, also known as Motility-related protein-1 (MRP-1). CD9 is a 228 amino acid, including an N-terminal initiator methionine ~24-27 kDa multipass transmembrane glycoprotein and member of the tetraspanin family. It is thought that the main role of tetraspanins is to form signal transducing complexes with integrins at the cell surface, for this reason CD9 has multiple functions in cell adhesion and motility, platelet activation, gamete fusion and cell proliferation.CD9 has a functional role as a tumour metastatic suppressor and is expressed in melanomas, and colon, lung and breast carcinomas.
The antibody reacts with mouse Mrp6, a 165 kD transmembrane protein that is related to the multidrug resistance related protein Mrp1. Mutations in the human MRP6 gene are responsible for the connective tissue disorder Pseudoxanthoma elasticum (PXE). M6II-24 was raised against a bacterial fusion protein of mouse Mrp6, containing amino acids 843-999
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
M6II-68
Concentration:
50 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
M6II-24 reacts with mouse Mrp6, a 165 kD transmembrane protein that is related to the multidrug resistance related protein Mrp1. Mutations in the human MRP6 gene are responsible for the connective tissue disorder Pseudoxanthoma elasticum (PXE). M6II-24 was raised against a bacterial fusion protein of mouse Mrp6, containing amino acids 843-999.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
M6II-24
Concentration:
50 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
This antibody reacts specifically with the MHC class II epitope present on Strain 13, but not on Strain 2 guinea pig cells. The antigen is also found in outbred Hartley guinea pigs.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
(CI.13.1)
Concentration:
700 ul/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
Mouse anti Guinea Pig CD90 antibody, clone CT4 recognizes guinea pig THY-1 (CD90), a small but heavily glycosylated member of the immunoglobulin superfamily. Clone CT4 was originally identified as recognizing a guinea pig homing receptor (Kraal et al. 1986) and later confirmed as recognizing the guinea pig homologue of human and rodent CD90 (Schäfer et al. 1999) by purification and microsequencing. Guinea Pig CD90 is notable for its level of N-linked glycosylation, rendering it an apparent molecular mass of ~36kDa, higher than that seen in most other species where CD90 has been identified and characterized. Guinea pig CD90 (according to Uniprot entry Q9WUR5) demonstrates 82% amino acid identity with human CD90, 74% with murine CD90 and 76% with rat.
Antibody Isotype:
IgG3
Monosan Range:
MONOSAN
Clone:
CT-4
Concentration:
500 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
The MDR Sampler Pack contains 4x 250ul Multidrug-resistance related protein antibodies: anti-MDR clone JSB-1 (MON9011P), anti-MDR clone LRP-56 (MON9016), anti-MDR clone MRPm6 (MON9017-1), anti-MDR clone MRPr1 (MON9018-1). Please see the specific datasheets for more information.
Monosan Range:
MONOSAN
Clone:
Sampler
Concentration:
100-250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
The gene coding for p53 oncoprotein is located on chromosome 17p. Allele loss at this chromosome site has frequently been seen in many tumours including lung, colon, breast and brain. Expression of p53 oncoprotein was not detected in normal mucosa, whereas mutation results in a detectable expression of the p53 protein. It is therefore suggested that immunocytochemical detection of p53 protein is an indication for malignancy. Positive control: Breast carcinoma.
p16 (CDKN2A, p16Ink4A), is key regulator of the cell cycle and involved in cell cycle control and cellular senescence. It is a specific inhibitor for Cdk4 and Cdk6 and binds to the phosphorylated Cdk-cyclin complex. A disruption of this pathway is commonly observed in cancer. p16 is lost in the majority of tumor cell lines and in most primary tumors. It is not expressed in melanoma.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
DCS-50
Concentration:
n/a
Storage buffer:
Lyophilized; reconstitute in 1 ml dist. water (final solution contains 0.09% sodium azide, 0.5% BSA in PBS buffer, pH 7.4)
Storage:
2-8°C
References 1:
Wiest, T. et al. Oncogene 2002;21: 15101517
References 2:
Shibata, K. R. et al. Stem Cells 2007; 25: 237182
NKX3.1 is a prostate specific androgen-regulated homeobox gene located on chromosome 8p. It is difficult to distinguish between high grade prostate adenocarcinoma and high grade infiltrating urothelial carcinoma using hematoxylin and eosin stained specimens. Current prostate adenocarcinoma markers such as prostate specific antigen (PSA) and prostate specific acid phosphatase (PSAP) are very useful in determining prostate origin of prostate cancer in other sites, but have lower sensitivity when identifying poorly differentiated compared to well differentiated cases. NKX3.1 is a sensitive and specific tissue marker of prostate adenocarcinoma and can be used to help distinguish it from urothelial carcinomas. Currently, thrombomodulin and uroplakin are used to identify tumors of urothelial origin; however, their sensitivities are suboptimal.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
EP356
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Gurel B, et al. Am J Surg Pathol. 2010; 34:1097-105
References 2:
Chuang AY, et al. Am J Surg Pathol. 2007; 31:1246-55
Immunoglobulin E (IgE) is a 180 kDa soluble protein serving as an antigen-specific unit of mast cell effector mechanisms. IgE has the lowest serum concentration of all immunoglobulins (approximately 0.5 mg/l) in healthy individuals, but upon allergen challenge its concentration in blood increases dramatically. Although biological survival of free IgE is very short (T1/2 = 2 days), it is stabilized after binding to its high affinity receptor. Unlike IgM- IgG- and IgA-committed B cells, IgE-switched B cells do not undergo clonal expansion.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
4H10
Concentration:
1 mg/ml
Format:
Purified by protein-A affinity chromatography.
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
M7I-3 reacts with an internal epitope of MRP7 (ABCC10), an approximately 160 kD transmembrane protein that is related to the multidrug resistance protein MRP1. M7I-3 was raised against a bacterial fusion protein of human MRP7, containing amino acids 194-272 of the protein.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
M7I-3
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
M5I-10 reacts with an internal epitope of MRP5 (ABCC5), an approximately 160 kD transmembrane protein that is related to the multidrug resistance protein MRP1. M5I-10 was raised against a bacterial fusion protein of mouse Mrp5, containing amino acids 1-38 of the protein. The Mab also strongly reacts with the human MRP5 protein.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
M5I-10
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
M9II-3 reacts with an internal epitope of MRP9 (ABCC12), an approximately 150 kD transmembrane protein that is related to the multidrug resistance protein MRP1. M9II-3 was raised against a bacterial fusion protein of human MRP9, containing amino acids 690-734 of the protein.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
M9II-3
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Ono N et al. Biochem J 2007; 406: 31-40
References 2:
Kruh GD et al. Pflugers Arch 2007; 453: 675-84
References 3:
Bera TK et al. Proc Natl Acad Sci 2002; 99: 6997-7002
M9I-38 reacts with an internal epitope of MRP9 (ABCC12), an approximately 150 kD transmembrane protein that is related to the multidrug resistance protein MRP1. M9I-38 was raised against a bacterial fusion protein of human MRP9, containing amino acids 1-42 of the protein.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
M9I-38
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Ono N et al. Biochem J 2007; 406: 31-40
References 2:
Kruh GD et al. Pflugers Arch 2007; 453: 675-84
References 3:
Bera TK et al. Proc Natl Acad Sci 2002; 99: 6997-7002
M8I-74 reacts with an internal epitope of MRP8 (ABCC11), an approximately 150 kD transmembrane protein that is related to the multidrug resistance protein MRP1. M8I-74 was raised against a bacterial fusion protein of human MRP8, containing amino acids 1-83 of the protein.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
M8I-74
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Kruh GD et al. Pflugers Arch 2007; 453: 675-84
References 2:
De Wolf CJ et al. FEBS J 2007; 274:439-50
References 3:
Oguri T et al. Mol Cancer Ther 2007; 6: 122-7
References 4:
Park S et al. Breast Cancer Res Treat 2006; 99: 9-17
The presence of prostate specific antigen is highly specific for demonstration of the prostatic origin of a malignancy. Mab ER-PR8 has been shown to be a valuable reagent for the identification of primary and metastatic prostatic carci-noma. The antibody reacts with a 34kD protein in benign and malignant prostatic epithelium. Positive control: Prostatic benign hyperplasia.
CD16 (FcgammaRIII) is a 50-65 kDa glycoprotein serving as a low affinity IgG receptor. Human FcgammaRIII is expressed in two forms – FcgammaRIII-A and -B. FcgammaRIII-A is a transmembrane protein of monocytes, macrophages, NK cells and a subset of T cells. It is associated with FcepsilonRI-gamma subunit and is responsible for antibody-dependent NK cell cytotoxicity. Mast cell FcgammaRIII-A is associated, moreover, with FcepsilonRI-beta subunit. Besides IgG, FcgammaRIII-A can be triggered also by oligomeric IgE. FcgammaRIII-B is a GPI-linked monomeric receptor expressed on neutrophils and is involved in their activation and induction of a proadhesive phenotype.
Immunoglobulin classes share the same basic four polypeptide chain structure of two heavy chains (five heavy chains types) and two light chains (kappa, lambda; both having a molecular weight of 22.5kDa). Kappa and lambda consist of a variable region and a constant region and can easily be differentiated by the antigenic properties of the constant region. The ratio of kappa to lambda is 70:30.
The human Mucin-1 antibody is reactive with 90% of breast carcinoma, most epithelial ovarian carcinoma and a high portion of the lung, urogenital and gastrointestinal carcinomas. Some reactivity is observed in the apical membrane of normal breast epithelium. Weak reactivity is also present in pancreas, kidney and lung. BC-2 reacts with a five amino acid epitope of the MUC-1 core protein which is less affective for glysosilation than most other MUC-1 epitopes. Localization: cytoplasm and membrane. Positive control: Mamma carcinoma.
CD54 (ICAM-1) is a 90 kD member of the C2 subset of immunoglobulin superfamily. It is a transmembrane molecule with 7 potential N-glycosylated sites, expressed on resting monocytes and endothelial cells and can be upregulated on many other cells, e.g. with lymphokines, on B- and T-lymphocytes, thymocytes, dendritic cells and also on keratinocytes, chondrocytes, as well as epithelial cells. CD54 mediates cell adhesion by binding to integrins CD11a/CD18 (LFA-1) and to CD11b/CD18 (Mac-1). The interaction of CD54 with LFA-1 enhances antigen-specific T-cell activation.
Oct-binding factor-1 (OBF1), also known as BOB.1, is a B-cell-specific coactivator which has been mapped to chromosome 11q23. Expression of BOB.1/OBF.1 is restricted largely to mature B-cells, with germinal center B-cells normally staining for BOB.1.2,3 Analyses of BOB.1/OBF.1 expression in a variety of established B-cell lines representing different stages of B-cell development has suggested a constitutive, B-cell-specific expression pattern. LP cells in nodular lymphocyte predominant Hodgkin lymphoma, because they are germinal center-derived, are consistently immunoreactive for BOB.1. Conversely, only some cases of classical Hodgkin lymphoma show BOB.1 immunoreactivity within the Hodgkin and Reed-Sternberg cells. Expression of BOB.1/OBF.1 has been reported in follicular center cell lymphoma, diffuse large B-cell lymphoma and some cases of acute myeloid leukemia. B-CLL, marginal zone lymphoma, and mantle cell lymphoma may show weak to moderate immunoreactivity.
Antibody Isotype:
IgG-1
Monosan Range:
MONOSAN
Clone:
SP92
Concentration:
n/a
Storage buffer:
Tris Buffer, pH 7.3-7.7, containing 1% BSA and <0.1% Sodium Azide
Storage:
2-8°C
References 1:
Junker S, et al. Genomics. 1996; 33:143-5
References 2:
Steimle-Grauer SA, et al. Virchows Arch. 2003; 442:284-93
License is required for use outside research field. Fibrinogen is a protein produced by the liver which helps stop bleeding by helping blood clots to form. Fibrinogen gets deiminated (conversion from arginin to citrullin) by Peptidyl Arginine Deïminase (PAD) in inflamed joints in patients that develop rheumatoid arthritis. Citrulline, while being an amino acid, is not built into proteins during protein synthesis, as it is not coded for by DNA, yet several proteins are known to contain citrulline. Proteins that normally contain citrulline residues include myelin basic protein (MBP), filaggrin, and several histone proteins, while other proteins, like fibrin and vimentin can get deiminated during cell death and tissue inflammation. Patients with rheumatoid arthritis often (at least 80% of them) develop an immune response against proteins containing citrulline. Although the origin of this immune response is not known, detection of antibodies reactive with citrulline containing proteins or peptides is now becoming an important help in the diagnosis of rheumatoid arthritis.
Chicken IgY is specific to chickens and is the counterpart to IgG from mammals. Chickens transfer high quantities of IgY into the egg yolk and harvesting antibodies from eggs eliminates the need for the invasive bleeding procedure. One weeks eggs can contain 10 times more antibodies than the volume of rabbit blood obtained from one weekly bleeding. Due to the phylogenetic distance between birds and mammals, there is greater potential of producing a higher percentage of specific antibody against mammalian antigens when using chickens [1]. Since chicken IgY does not cross-react with mammalian IgG [2] and does not bind bacterial or mammalian Fc receptors [3], non-specific binding is reduced, and the need for cross-species immunoabsorptions is also eliminated [4].
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
08C
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium
Storage:
2-8°C
References 1:
Jensenius et al. J. Immunol Meth 1981; 46:63
References 2:
Ambrosius et al. Vet Immunol. Immunopathol;17:57
References 3:
Larsson et al, J. Immunol Meth 1988; 108:205
References 4:
Larsson et al. Comp. Immun.Microbiol Infe. Dis 1990; 13:199
License is required for use outside research field. Fibrinogen is a protein produced by the liver which helps stop bleeding by helping blood clots to form. Fibrinogen gets deiminated (conversion from arginin to citrullin) by Peptidyl Arginine Deïminase (PAD) in inflamed joints in patients that develop rheumatoid arthritis. Citrulline, while being an amino acid, is not built into proteins during protein synthesis, as it is not coded for by DNA, yet several proteins are known to contain citrulline. Proteins that normally contain citrulline residues include myelin basic protein (MBP), filaggrin, and several histone proteins, while other proteins, like fibrin and vimentin can get deiminated during cell death and tissue inflammation. Patients with rheumatoid arthritis often (at least 80% of them) develop an immune response against proteins containing citrulline. Although the origin of this immune response is not known, detection of antibodies reactive with citrulline containing proteins or peptides is now becoming an important help in the diagnosis of rheumatoid arthritis.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
23H2
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
License is required for use outside research field. Fibrinogen is a protein produced by the liver which helps stop bleeding by helping blood clots to form. Fibrinogen gets deiminated (conversion from arginin to citrullin) by Peptidyl Arginine Deïminase (PAD) in inflamed joints in patients that develop rheumatoid arthritis. Citrulline, while being an amino acid, is not built into proteins during protein synthesis, as it is not coded for by DNA, yet several proteins are known to contain citrulline. Proteins that normally contain citrulline residues include myelin basic protein (MBP), filaggrin, and several histone proteins, while other proteins, like fibrin and vimentin can get deiminated during cell death and tissue inflammation. Patients with rheumatoid arthritis often (at least 80% of them) develop an immune response against proteins containing citrulline. Although the origin of this immune response is not known, detection of antibodies reactive with citrulline containing proteins or peptides is now becoming an important help in the diagnosis of rheumatoid arthritis.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
20B2
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
License is required for use outside research field. Fibrinogen is a protein produced by the liver which helps stop bleeding by helping blood clots to form. Fibrinogen gets deiminated (conversion from arginin to citrullin) by Peptidyl Arginine Deïminase (PAD) in inflamed joints in patients that develop rheumatoid arthritis. Citrulline, while being an amino acid, is not built into proteins during protein synthesis, as it is not coded for by DNA, yet several proteins are known to contain citrulline. Proteins that normally contain citrulline residues include myelin basic protein (MBP), filaggrin, and several histone proteins, while other proteins, like fibrin and vimentin can get deiminated during cell death and tissue inflammation. Patients with rheumatoid arthritis often (at least 80% of them) develop an immune response against proteins containing citrulline. Although the origin of this immune response is not known, detection of antibodies reactive with citrulline containing proteins or peptides is now becoming an important help in the diagnosis of rheumatoid arthritis.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
3D1
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
Protein tyrosine phosphatases (PTPs) are the enzymes that are instrumental in determining the spatial and temporal balance between the tyrosine phosphorylated and non-phosphorylated targets, and thus coordinately regulate these cellular responses to extracellular cues. The Ptprr gene gives rise to 4 different neuronal phosphatases which differ in length of their Nterminal part and subcellular localization. PTPBR7 (72 kDa) is receptor-like, PTP-SL (60 kDa) is membrane associated and PTPPBS? (42 and 37 kDa) is a cytosolic phoaphatase. This antibody is directed to the common part of these proteins.
Antibody Isotype:
IgG2
Monosan Range:
MONOSAN
Clone:
6A6
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
MUC4 (mucin 4) is a large membrane-anchored glycoprotein of the mucin family that is expressed by epithelial cells in various normal tissues including lung, bronchus, stomach, colon, and cervix. MUC4 is generally not detected in normal pancreas, but is expressed in the vast majority of pancreatic neoplasms, such as pancreatic ductal adenocarcinoma. Additionally, expression in various carcinomas has been reported, including gastric adenocarcinoma, colon adenocarcinoma, and lung adenocarcinoma.
Fluorescent proteins, like EBFP, can be used as protein "tags" to study the subcellular localization of proteins and/or their translocation upon stimulation and/or as markers for transfections in transient and stable expression systems. In immunoblot it cross reacts with GFP and YFP.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
7F10
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
Fluorescent proteins, like EBFP, can be used as protein "tags" to study the subcellular localization of proteins and/or their translocation upon stimulation and/or as markers for transfections in transient and stable expression systems. In immunoblot it cross reacts with GFP and YFP.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
3A6
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
The exosome, present in both the nucleus and cytoplasm of all eukaryotic cells, is a complex of 3'-5' exoribonucleases containing at least nine core components. Recently, it has been demonstrated, mainly by analyses in yeast, that the nuclear exosome is essential for rRNA processing and sn(o)RNA biogenesis. Furthermore, it is involved in the degradation of improperly processed mRNAs. The cytoplasmic exosome participates in normal mRNA turnover and in the degradation of inherently instable mRNAs that contain AU-rich elements. Therefore, the exosome plays a key role in RNA metabolism.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
15B3
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
The nuclei of eukaryotic cells contain several classes of small RNA-protein complexes. SnRNPs (small nuclear ribonucleoproteins) are involved in splicing of pre-mRNAs (the excision of introns) in the nucleoplasm. RPP20 is part of such a complex. SnoRNPs (small nucleolar ribonucleoproteins) are involved in the maturation of pre-ribosomal RNA in the nucleoli. Pre-rRNA processing and modification require both the snoRNA and the protein constituents (such as Rpp20) of these complexes. Generally, snoRNPs contain a number of common proteins, which are shared with other snoRNPs of the same family, next to several particle-specific proteins. On average, snoRNPs contain about 6-10 protein subunits
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
1F11
Concentration:
n/a
Storage buffer:
lyophilized in a 10 mM ammonium bicarbonate buffer. Each vial contains 2 mg BSA
CD138 (syndecan 1) is a transmembrane proteoglycan that can bind a variety of cytokines and modulate their activity, as well as the activity of extracellular matrix components and influence many developmental processes. CD138 is expressed mainly in differentiating keratinocytes and is transiently upregulated in all layers of the epidermis upon tissue injury. It is also highly expressed on plasma cells and can be detected even on fibroblasts, vascular smooth muscle cells and endothelial cells. Up-regulation and down-regulation of CD138 on the cell surface often correlates with the gain of cancerous characteristics. Serum levels of the shedded soluble sCD138 are used as a prognostic factor of cancerogenesis.Purified by protein-A affinity chromatography. The mouse monoclonal antibody MI15 recognizes an extracellular epitope of CD138 (syndecan 1), a 65-70 kDa heparan sulfate proteoglycan expressed mainly in the epidermis and plasma cells, but also in growth factor-stimulated lymphocytes.
Antibody Isotype:
IgG1 kappa
Monosan Range:
MONOSAN
Clone:
MI15
Concentration:
1 mg/ml
Storage buffer:
PBS pH 7.4, 15 mM sodium azide
Storage:
2-8°C
References 1:
Nadalin MR et al. Braz Dent J. 2011;22(3):223-9
References 2:
Noll JE et al. J Hematol Oncol. 2015 Oct 6;8:106
References 3:
Krishnan SR et al. Neoplasia. 2016 Jan;18(1):25-32
References 4:
Jourdan M et al. J Immunol. 2011 Oct 15;187(8):3931-41
References 5:
Atanackovic D et al. J Natl Cancer Inst. 2012 Jul 3;104(13):1005-20
HLA-class I major histocompatibility (MHC) antigens are intrinsic membrane glycoproteins expressed on nucleated cells and noncovalently associated with an invariant beta2 microglobulin. They carry foreign determinants important for immune recognition by cytotoxic T cells, thus important for anti-viral and anti-tumour defence. Classical human HLA-class I antigens are represented by HLA-A, HLA-B and HLA-C molecules, the non-classical by e.g. HLA-E, HLA-G.
HLA-class I major histocompatibility (MHC) antigens are intrinsic membrane glycoproteins expressed on nucleated cells and noncovalently associated with an invariant beta2 microglobulin. They carry foreign determinants important for immune recognition by cytotoxic T cells, thus important for anti-viral and anti-tumour defence. Classical human HLA-class I antigens are represented by HLA-A, HLA-B and HLA-C molecules, the non-classical by e.g. HLA-E, HLA-G.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
TP25.99SF
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The antibody reacts with 170 - 220 kD cell surface glycoproteins (CD45), the Leukocyte Common Antigen (LCA), expressed selectively on hematopoietic cells. It gives a pronounced staining on formalin-fixed, paraffin-embedded cells, and can be used to differentiate between lymphomas and carcinomas. Positive control: Tonsil.
CD30 is a type I transmembrane glycoprotein of the TNF receptor superfamily. CD30 was originally identified as a cell surface antigen of Hodgkins and Reed-Sternberg cells using monoclonal antibody Ki-1. The ligand for CD30 is CD30L (CD153). The binding of CD30 to CD30L mediates pleiotropic effects including cell proliferation, activation, differentiation, and apoptotic cell death. CD30 has a critical role in the pathophysiology of Hodgkin's disease and other CD30+ lymphomas. CD30 acts as a costimulatory molecule in thymic negative selection. In addition to its expression on Hodgkin's and Reed-Sternberg cells, CD30 is also found in some non-Hodgkin's lymphomas (including Burkitt's lymphomas), virus-infected T and B cells, and on normal T and B cells after activation. In T cells, CD30 expression is present on a subset of T cells that produce Th2-type cytokines and on CD4+/CD8+ thymocytes that co-express CD45RO and the IL4 receptor. Soluble form of CD30 (sCD30) serves as a marker reflecting Th2 immune response.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
MEM-268
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
HLA-DR1 belongs to the HLA class II beta chain paralogues. The MHC Class II molecule is a heterodimer consisting of an alpha (DRA) and a beta chain (DRB), both anchored in the membrane. It plays a central role in the immune system by presenting peptides derived from extracellular proteins. MHC Class II molecules are expressed in antigen presenting cells (APC). The beta chain is approximately 26-28 kDa. Within the DR molecule the beta chain contains all the polymorphisms specifying the peptide binding specificities. Hundreds of DRB1 alleles have been described and typing for these polymorphisms is routinely done for bone marrow and kidney transplantation.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
MEM-267
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
TRAIL-R2 (CD262, DR5) is one of two TNF superfamily member intracellular death domain containing receptors for TRAIL (APO2L). Apoptosis, or programmed cell death, occurs during normal cellular differentiation and development of multicellular organisms. Apoptosis is induced by certain cytokines including tumor necrosis factor (TNF) and Fas ligand in the TNF family through their death domain containing receptors, TNF receptor 1 (TNFR1) and Fas, respectively. Another member in the TNF family has been identified and designated TRAIL (for TNF related apoptosis inducing ligand) and Apo2L (for Apo2 ligand). Receptors for TRAIL include two death domain containing receptors, DR4 and DR5, as well as two decoy receptors, DcR1 and DcR2, lacking the intracellular signaling death domain. DcR1 (also called TRID), like the related death receptors DR4 and DR5, contains two extracellular cysteine rich domains. However, DcR1 contains no intracellular death domain and is thus incapable of signaling apoptosis. It has been suggested DcR1 is responsible for TRAIL resistance in normal human tissues including heart, placenta, lung, liver, kidney, spleen, and bone marrow. DR5 is a member of the TNF receptor superfamily, and contains an intracellular death domain. This receptor can be activated by tumor necrosis factor related apoptosis inducing ligand (TNFSF10/TRAIL/APO2L), and transduces apoptosis signal. Studies with FADD deficient mice suggested that FADD, a death domain containing adaptor protein, is required for the apoptosis mediated by this protein.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
DR5-01
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The M4I-80 Mab was selected after immunization with a fusion protein containing the E. coli maltose binding protein and a fragment of the human MRP4 protein corresponding to amino acids 372-431. MRP4 transports cyclic nucleotides and anti-retroviral compounds. The M4I-80 Mab also reacts with the mouse orthologue of the transporter molecule (Mrp4).
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
M4I-80,
Concentration:
200 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
The antibody reacts with free soluble (17 kDa) and membrane (26 kDa) human TNF-alpha. The antibody inhibits the biological activity of both forms. It does not react with receptor bound TNF-alpha. It can be a useful tool to discriminate between the membrane form of TNF expressed on producer cells and the proteolytically cleaved, soluble TNF-alpha bound to its cognate cell membrane receptors (TNF-RI and TNF-RII). For this purpose we recommend to use this antibody in combination with the anti-TNF-alpha antibody HM2026, which recognizes soluble, membrane and receptor bound TNF-alpha.
The antibody reacts with free soluble (17 kDa) and membrane (26 kDa) human TNF-alpha. The antibody inhibits the biological activity of soluble and membrane TNF-alpha. The antibody can be a useful tool to discriminate between receptor bound soluble (17 kDa) and the membrane (26 kDa) form of TNF-alpha. For this purpose we recommend to use this antibody in combination with the anti-TNF-alpha antibody HM2024, which recognizes only soluble and membrane TNF-alpha, but not the receptor bound TNF-alpha.
The monoclonal antibody HM102 recognizes the extracellular part of membrane-bound TNF-RII as well as the soluble form of TNF-RII which is generated by proteolytic cleavage of the extracellular domain. The soluble form can still bind TNF-alpha with high affinity and functions as a TNF-alpha antagonist. TNF-alpha is an important signalling protein in the immune system which can activate inflammatory responses, induce apoptosis, regulate cellular proliferation, and may even promote cancer progression. TNF-alpha can bind to two structurally distinct membrane receptors, TNF-RI and TNFRII, which have both distinct and overlapping downstream signaling cascades. TNFRI is believed to be expressed on nearly all cell types, whereas TNFRII exhibits more restricted expression, being found on certain subpopulations of immune cells and several other cell types. A dominant role of TNFRII has been shown in thymocyte activation by TNF-alpha, whereas induction of cytotoxicity and other functions are mediated largely by TNF-RI. TNF-RI is equally well activated by both the 17 kDa soluble and 26 kDa membrane-bound form, whereas TNF-RII is activated only by the membrane bound form of TNF-alpha. The antibody is a agonistic receptor modulating antibody. It enhances in vitro TNF alpha responses by increasing the affinity of the soluble form of TNF-alpha for TNF-RII.
The monoclonal antibody HM104 recognizes the extracellular part of the Tumor Necrosis Factor Receptor type I (TNF-RI) of the membrane-bound as well as the soluble receptor. TNF-RI (~55-60 kDa) is present on most cell types and is considered to play a prominent role in cell stimulation by TNF-alpha. TNF-alpha activates inflammatory responses, induces apoptosis, regulates cellular proliferation, and may even promote cancer progression. The effects of TNF-alpha are mediated by TNF-RI and TNF-RII, which have both distinct and overlapping downstream signaling cascades. Induction of cytotoxicity and other functions are mediated largely via TNF-RI. TNF-RI is equally well activated by both the 17 kDa soluble and 26 kDa membrane-bound form, whereas TNF-RII is efficiently activated only by the membrane bound form of TNF-alpha. TNF-RI signaling is initiated when trimeric TNF-alpha binds TNF-RI receptors. Subsequent TNF-RI trimerization promotes the recruitment of a proximal signaling complex composed of TNF Receptor Associated protein with a Death Domain (TRADD), Receptor Interacting Protein (RIP), cellular Inhibitor of Apoptosis Protein 1 (cIAP1), TNF Receptor Associated Factor 2 (TRAF2), and likely TRAF5. Studies with TNF-RI-deficient mice indicate that TNF-RI mediates most of the proliferation, pro-inflammatory, and apoptosis-activating pathways.
The antibody MR2-1 reacts with the extra-cellular part of the TNF-RII. It also reacts with the soluble receptor. TNF-RII is present on most cell types and is considered to play a prominent role in cell stimulation by TNF-alpha. TNF-RII molecule is shown to be responsible for stimulation of activated T-lymphocytes by TNF-alpha. The antibody cross reacts with rhesus and cynomolgus natural TNF-RII.
The antibody MR1-2 reacts with the extra-cellular part of the TNF-RI. It also reacts with the soluble receptor. TNF-RI is present on most cell types and is considered to play a prominent role in cell stimulation by TNF-alpha: Induction of cytotoxicity and other functions are mediated largely via TNF-RI. The antibody cross reacts with rhesus and cynomolgus natural TNF-RI.
The antibody reacts with human native and recombinant TNF-alpha as assessed by ELISA. The antibody inhibits the biological activity of human native and recombinant TNF-alpha as determined with L929 cells in a cytotoxicity assay. The antibody cross reacts with rhesus and cynomolgus natural TNF-alpha and lacks crossreactivity with human lymphotoxin.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
4H31
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Gerspach; J et al. Microsc Res Tech 2000; 50: 243
References 2:
Limb, GA et al Br J Ophthalmol 1996, 80: 168
References 3:
Laan van der; N et al. Arch Dermatol Res 2001; 293: 226
The monoclonal antibody V1q recognizes mouse tumor necrosis factor alpha (TNF-?). TNF-? is the prototype cytokine of the family of TNF-related ligands, which are based on structural and functional homologies. TNF-? is synthesized as type II transmembrane protein. TNF-? can be recognized by two different membrane receptors, namely TNF-R1 and TNF-R2. TNF-? is present in a membrane-bound (tmTNF) as well as soluble form (sTNF). The membrane-bound form of TNF-? is recognized by both TNF receptors with high affinity, whereas the soluble form is recognized more superiorly by TNF-R1. TNF-? is produced by many different cell types including macrophages, T lymphocytes, NK cells, neutrophils and endothelial cells. Cells differ in the expression of the two TNF-receptors and sTNF versus tmTNF, respectively. TNF-?, a homotrimeric 17 kDa protein, is a potent mediator of inflammatory and metabolic functions. TNF-? was originally detected as a highly cytotoxic cytokine for tumor cells, it causes tumor necrosis in vivo and shows cytolytic activity against tumor cells in vitro. Furthermore, TNF-? has been implied as central mediator in shock induced by gram negative micro-organisms. TNF-? induces on its turn the production of many other cytokines. Furthermore, TNF-? has been found in inflammatory foci such as synovial effusions in rheumatoid arthritis, systemic circulation in septic shock, parasitemia and rejection of renal transplants. The monoclonal antibody V1q recognizes both natural and recombinant TNF-? and shows neutralizing activity.
Monosan Range:
MONOSAN
Clone:
V1q
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Echtenacher; B et al. J Immunol 1990; 145: 3762
References 2:
Gerspach, J et al Microsc Res Tech 2000, 50: 243
References 3:
Demjen; D et al. Nat Med 2004; 10: 389
References 4:
Rajashekhar G et al. Physiol Genomics 2007; 31: 104
The monoclonal antibody 80M2 recognizes the extracellular part of- membrane-bound TNF-RII as well as- the soluble form of TNF-RII which is generated by proteolytic cleavage of the extracellular domain. The soluble form can still bind TNF-alpha with high affinity and functions as a TNF-alpha antagonist.- TNF-alpha is an important signaling protein in the immune system which can activate inflammatory responses, induce apoptosis, regulate cellular proliferation, and may even promote cancer progression. TNF-alpha can bind to two structurally distinct membrane receptors, TNF-RI and TNF-RII, which have both distinct and overlapping downstream signaling cascades. TNFRI is believed to be expressed on nearly all cell types, whereas TNFRII exhibits more restricted expression, being found on certain subpopulations of immune cells and several other cell types. A dominant role of TNF-RII has been shown in thymocyte activation by TNF-alpha, whereas induction of cytotoxicity and other functions are mediated largely by TNF-RI. TNF-RI is equally well activated by both the 17 kDa soluble and 26 kDa membrane-bound form, whereas TNF-RII is activated only by the membrane bound form of TNF-alpha. The antibody is a non-agonistic receptor modulating antibody. It enhances in vitro TNF alpha responses by increasing the affinity of the soluble form of TNF-alpha for TNF-RII.
The monoclonal antibody H398 recognizes the extracellular part of the Tumor Necrosis Factor Receptor type I (TNF-RI) of the membrane-bound as well as the soluble receptor. TNF-RI (~55-60 kDa) is present on most cell types and is considered to play a prominent role in cell stimulation by TNF-alpha. TNF-alpha activates inflammatory responses, induces apoptosis, regulates cellular proliferation, and may even promote cancer progression. The effects of TNF-alpha are mediated by TNF-RI and TNF-RII, which have both distinct and overlapping downstream signaling cascades. Induction of cytotoxicity and other functions are mediated largely via TNF-RI. TNF-RI is equally well activated by both the 17 kDa soluble and 26 kDa membrane-bound form, whereas TNF-RII is efficiently activated only by the membrane bound form of TNF-alpha. TNF-RI signaling is initiated when trimeric TNF-alpha binds TNF-RI receptors. Subsequent TNF-RI trimerization promotes the recruitment of a proximal signaling complex composed of TNF Receptor Associated protein with a Death Domain (TRADD), Receptor Interacting Protein (RIP), cellular Inhibitor of Apoptosis Protein 1 (cIAP1), TNF Receptor Associated Factor 2 (TRAF2), and likely TRAF5. Studies with TNF-RI-deficient mice indicate that TNF-RI mediates most of the proliferation, pro-inflammatory, and apoptosis-activating pathways.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
H398
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Thoma; B et al. J Exp Med 1990; 172: 1019
References 2:
Grell, M et al Lymphokine Cytokine Res 1993, 12: 143
References 3:
Scheurich; P et al. Tumor Necrosis factor 1993; 4: 52
References 4:
Grell M et al. Proc Natl Acad Sci USA 1998; 95: 570
References 5:
Krippner-Heidenreich A et al. J Immunol 2008; 180: 8176
MRP4 transports cyclic nucleotides and anti-retroviral compounds. The M4I-10 Mab also reacts with the mouse orthologue of the transporter molecule (Mrp4).
The antibody reacts with an internal epitope of MRP2, a 190-200 kD transmembrane protein earlier known as the canalicular multi-organic anion transporter cMOAT, absent in patients with the Dubin-Johnson syndrome, an autosomal recessive liver disorder characterized by chronic conjugated hyperbilirubinemia. MRP2 is a member of the MRP family of multidrug resistance related proteins, and MRP2 overexpression has been observed in a subset of cisplatin resistant cell lines. M2III-5 was raised against a fusion protein of the bacterial maltose binding protein and rat Mrp2, containing the 202-amino acid COOH terminal end of the transporter protein. The Mab detects rat, mouse and human MRP2. M2III-5 does not cross-react with the human MDR1 P-gp, MRP1, MRP3 orMRP5 gene products.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
M2III-5
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
The rat monoclonal antibody LMR-42 detects an outer-surface epitope of a 55 kDa plasma membrane protein overexpressed in several human non-Pgpmultidrug-resistant (MDR) tumor cell lines. Expression cloning revealed that the LMR-42 antigen is identical to the human endothelial cell protein Creceptor (EPCR). Protein C is the zymogen of the key anticoagulant enzyme, activated protein C (APC).
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
LMR-42
Concentration:
100 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Flens MJ et al. Int J Cancer 1997; 73: 249
References 2:
Scheffer GL et al. J Cancer 2002; 1535-42
References 3:
Esmon CT et al. Crit Care Med 2004; 5 suppl: s298-301
CD13 (aminopeptidase N, APN) is a 150 kDa type II transmembrane zinc-binding ectopeptidase expressed on various cell types. This metalloprotease preferentially catalyzes removal of neutral amino acids from small peptides, thus activating or inactivating bioactive peptides. CD13 has also role in extracellular matrix degradation, antigen processing and signal transduction, is important in inflammatory responses, regulates intercellular contact, cell motility and vascularization. CD13 is involved in protection of leukemic cells against apoptosis and its expression associated with poor prognosis of carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
WM15
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
BXP-53 reacts with an internal epitope of bcrp, a 70 kD transmembrane half-transporter which is involved in Multidrug resistance. BXP-53 also reacts with the human BCRP molecule.
The BXP-9 Mab was selected after immunization with a fusion protein containing the E. coli maltose binding protein and a fragment of the mouse bcrp protein corresponding to amino acids 221-394. BXP-9 reacts with an internal epitope of bcrp, a 70 kD transmembrane half-transporter which is involved in Multidrug resistance. BXP-9 does not react with the human BCRP molecule.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
BXP-9
Concentration:
250 ug/ ml
Storage buffer:
cell culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Doyle LA et al. Proc Nat Acad Sci 1998; 95: 15665-15670
M6II-31 reacts with MRP6, a 190-200 kD transmembrane protein that is related to the multidrug resistance related protein MRP. Mutations in the MRP6 gene are responsible for the connective tissue disorder Pseudoxanthoma elasticum (PXE). M6II-31 was raised against a bacterial fusion protein of human MRP6, containing amino acids 764-964, spanning the putative 12th transmembrane region as well as predicted internal and external regions of the protein. M6II-31 did not cross-react with the human MDR1, MRP1, MRP2, MRP3, MRP4, MRP5 gene products.
M6II-21 reacts with MRP6, a 190-200 kD transmembrane protein that is related to the multidrug resistance related protein MRP. Mutations in the MRP6 gene are responsible for the connective tissue disorder Pseudoxanthoma elasticum (PXE). M6II-21 was raised against a bacterial fusion protein of human MRP6, containing amino acids 764-964, spanning the putative 12th transmembrane region as well as predicted internal and external regions of the protein. M6II-21 did not cross-react with the human MDR1, MRP1, MRP2, MRP3, MRP4, MRP5 gene products. Unreactive on standard IHC-P.
M6II-7 reacts with MRP6, a 190-200 kD transmembrane protein that is related to the multidrug resistance related protein MRP. Mutations in the MRP6 gene are responsible for the connective tissue disorder Pseudoxanthoma elasticum (PXE). M6II-7 was raised against a bacterial fusion protein of human MRP6, containing amino acids 764-964, spanning the putative 12th transmembrane region as well as predicted internal and external regions of the protein. M6II-7 did not cross-react with the human MDR1, MRP1, MRP2, MRP3, MRP4, MRP5 gene products. unreactive on standard IHC-P.
1032 Is specific for the major vault protein, a 104-kDa highly conserved protein interacting with estrogen receptor. It is one of a series of four mAbs which recognize different epitopes of the protein. Major vault proteins have a complex morphology, including several small molecules of RNA, but a single protein species. The MVP accounts for >70% of their mass. Their shape is reminiscent of the nucleopore central plug. Treatment of cells with estradiol increases the amount of MVP in nuclear extract. The hormone-dependent interaction of vaults with ER is prevented in vitro by sodium molybdate. Antibodies to estrogen, progesterone and glucocorticoid receptors are able to co-immunoprecipitate the MVP. MVP is overexpressed in many neoplastic tissues and cell lines. Expression of MVP predicts a poor response to chemotherapy.
Antibody Isotype:
IgM
Monosan Range:
MONOSAN
Clone:
1032
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Abbondanza, C. et al, J. Cell Biol. 141, 1301-1310 (1998)
1027 Is specific for the major vault protein, a 104-kDa highly conserved protein interacting with estrogen receptor. It is one of a series of four mAbs which recognize different epitopes of the protein. Major vault proteins have a complex morphology, including several small molecules of RNA, but a single protein species. The MVP accounts for >70% of their mass. Their shape is reminiscent of the nucleopore central plug. Treatment of cells with estradiol increases the amount of MVP in nuclear extract. The hormone-dependent interaction of vaults with ER is prevented in vitro by sodium molybdate. Antibodies to estrogen, progesterone and glucocorticoid receptors are able to co-immunoprecipitate the MVP. MVP is overexpressed in many neoplastic tissues and cell lines. Expression of MVP predicts a poor response to chemotherapy.
Antibody Isotype:
IgM
Monosan Range:
MONOSAN
Clone:
1027
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Abbondanza, C. et al, J. Cell Biol. 141, 1301-1310 (1998)
1014 Is specific for the major vault protein, a 104-kDa highly conserved protein interacting with estrogen receptor. It is one of a series of four mAbs which recognize different epitopes of the protein. Major vault proteins have a complex morphology, including several small molecules of RNA, but a single protein species. The MVP accounts for >70% of their mass. Their shape is reminiscent of the nucleopore central plug. Treatment of cells with estradiol increases the amount of MVP in nuclear extract. The hormone-dependent interaction of vaults with ER is prevented in vitro by sodium molybdate. Antibodies to estrogen, progesterone and glucocorticoid receptors are able to co-immunoprecipitate the MVP. MVP is overexpressed in many neoplastic tissues and cell lines. Expression of MVP predicts a poor response to chemotherapy
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
1014
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Abbondanza, C. et al, J. Cell Biol. 141, 1301-1310 (1998)
1011 Is specific for the major vault protein, a 104-kDa highly conserved protein interacting with estrogen receptor. It is one of a series of four mAbs which recognize different epitopes of the protein. Major vault proteins have a complex morphology, including several small molecules of RNA, but a single protein species. The MVP accounts for >70% of their mass. Their shape is reminiscent of the nucleopore central plug. Treatment of cells with estradiol increases the amount of MVP in nuclear extract. The hormone-dependent interaction of vaults with ER is prevented in vitro by sodium molybdate. Antibodies to estrogen, progesterone and glucocorticoid receptors are able to co-immunoprecipitate the MVP. MVP is overexpressed in many neoplastic tissues and cell lines. Expression of MVP predicts a poor response to chemotherapy.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
1011
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Abbondanza, C. et al, J. Cell Biol. 141, 1301-1310 (1998)
47-8D3 reacts with macrophages and detects the well-known leukocyte L1, cystic fibrosis antigen. Detecting a single protein band of 14 kDa in Western blots of lysates of human monocytes and granulocytes, the antigen was identified as the calcium-binding protein MRP14, which is a member of the S100 family involved a.o. in regulating the cell cycle. MRP14 is also implicated in the abnormal differentiation of myeloid cells in the stroma of cancer. It is further found on squamous mucosal epithelia. When associated with MRP8 it forms the heterodimer calprotectin.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
47-8D3
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Flavell DJ. et al., J. Histochem. Cytochem. 35: 1217-1226 (1987)
References 2:
Facchetti F. et al., Am. J. Clin. Pathol. 92: 42-50 (1989)
References 3:
Bardadin KA. et al., J. Pathol. 164: 253-259 (1991)
References 4:
Goebeler M. et al., J. Leukocyte Biol. 55: 259-261 (1994)
BXP-21 reacts with an internal epitope of BCRP, a 70 kD transmembrane half-transporter which is involved in Multidrug resistance. BXP-21 did not cross-react with the human MDR1, MRP1, MRP2 gene products.
p193-6 reacts with an internal epitope (amino acids 593-599, FSKVEDY) of the minor vault protein (p193 or VPARP), which is overexpressed in various human non-P-glycoprotein MDR tumor cell lines, accordingly to an increase in the number of vault particles. p193-6 was raised against an E. coli lysate transformed with the pET28a(+) expression vector containing amino acids 408-611 of the p193 cDNA
p193-4 reacts with an internal epitope (amino acids 491-494, HPGE) of the minor vault protein (p193 or VPARP), which is overexpressed in various human non-P-glycoprotein MDR tumor cell lines, accordingly to an increase in the number of vault particles. p193-4 was raised against an E. coli lysate transformed with the pET28a(+) expression vector containing amino acids 408-611 of the p193 cDNA.
p193-10 reacts with an internal epitope (amino acids 506-510, VALGK) of the minor vault protein (p193 or VPARP), which is overexpressed in various human non-P-glycoprotein MDR tumor cell lines, accordingly to an increase in the number of vault particles. p193-10 was raised against an E. coli lysate transformed with the pET28a(+) expression vector containing amino acids 408-611 of the p193 cDNA.
BXP-34 Mab was selected after immunization with the mitoxanthrone resistant, BCRP overexpressing cell line MCF7 MR. BXP-34 reacts with an internal epitope of BCRP, a 70 kD transmembrane half-transporter which is involved in multidrug resistance. BXP-34 did not cross-react with the human MDR1, MRP1, MRP2, MRP5 gene products.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
BXP-34
Concentration:
250 ug/ ml
Storage buffer:
PBS with 1% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Doyle LA et al. Proc Nat Acad Sci 1998; 95: 15665-15670
References 2:
Scheffer GL et al. Cancer Res 2000; 60: 2589-2593
References 3:
van der Kolk DM et al. Blood; 99: 3763-3770
References 4:
van der Pol MA et al. Haematologica 2003; 88: 134-147
MVP-37 reacts with an internal epitope of MVP/LRP (p110), which is strongly overexpressed in various human non-P-glycoprotein MDR tumor cell lines, accordingly to an increase in the number of vault particles.
The antibody was selected after immunization with a fusion protein consisting of Gluthathione S Transferase and a fragment of MDR3 P-gp comprising amino acid 629 - 692. P3II-26 reacts with an internal epitope of MDR3 P-gp. P3II-26 does not cross-react with the human MDR1 P-gp.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
P3II-26
Concentration:
100 ug/ ml
Format:
Protein G purified
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Scheffer G et al. Cancer Res 2000; 60: 5269-5277
References 2:
Hooiveld GJ et al. Gastroenterology 1999; 117; 678-687
M5II-54 reacts with an internal epitope of MRP5, a 190-200 kD transmembrane protein that is closely related to the multidrug resistance protein MRP. M5II-54 was raised against a bacterial fusion protein of MRP5, containing amino acids 722-910 of the protein. M5I-1 does not cross-react with the human MDR1, MRP1, MRP2 or MRP3 gene products
M5I-1 reacts with an internal epitope of MRP5, a 190-200 kD transmembrane protein that is closely related to the multidrug resistance protein MRP. M5I-1 was raised against a bacterial fusion protein of MRP5, containing amino acids 82-168 of the protein. M5I-1 does not cross-react with the human MDR1, MRP1, MRP2 or MRP3 gene products
M2II-12 reacts with an internal epitope of cMOAT/MRP2, a 190-200 kD transmembrane protein known as the canalicular multi-organic anion transporter, absent in patients with the Dubin-Johnson syndrome, an autosomal recessive liver disorder characterized by chronic conjugated hyperbilirubinemia. cMOAT/MRP2 is closely related to the multidrug resistance related protein MRP, and cMOAT/MRP2 overexpression has been observed in a subset of cisplatin resistant cell lines. M2II-12 was raised against a bacterial fusion protein of cMOAB/M¬RP2, containing amino acids 860-950 of the protein. M2II-12 did not cross react with the human MDR1, MRP1, MRP3 and MRP5 gene products.
The antibody reacts with an internal epitope of MRP3, a 190-200 kD transmembrane protein that is closely related to the multidrug resistance protein MRP1. M3II-21 was raised against a bacterial fusion protein of MRP3, containing amino acids 830-949 of the protein. M3II-21 does not cross-react with the human MDR1, MRP1, MRP2 or MRP5 gene products. M3II-21 has potential value for detection of MRP3-mediated drug-resistance in human tumor samples.
M3II-9 reacts with an internal epitope of MRP3, a 190-200 kD transmembrane protein that is closely related to the multidrug resistance protein MRP1. M3II-9 was raised against a bacterial fusion protein of MRP3, containing amino acids 830-949 of the protein. M3II-9 does not cross-react with the human MDR1, MRP1, MRP2 or MRP5 gene products
The monoclonal antibody HM.11 recognizes modified amino acid nitrotyrosine in all different species. Nitrotyrosine is formed in tissues in presence of the active metabolite NO and is a stable end product of nitrosylation of tyrosine. Inflammation is characterized by increased nitric oxide (NO) production. NO reacts rapidly with superoxide to form peroxynitrite. At physiological pH and in the presence of transition metals, peroxynitrite undergoes heterolytic cleavage to form hydroxyl anion and nitronium ion, the latter of which nitrates protein tyrosine residues. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic plaques, Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS). Thus, the presence of nitrotyrosine on proteins can be used as a marker for peroxynitrite formation in vivo and consequently as a marker of NO-mediated tissue damage. The monoclonal antibody HM.11 recognizes nitrotyrosine, both with the free amino acid as well as with proteins containing nitrotyrosine
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
HM11
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Ter Steege; J et al. Free Radic Biol Med 1998; 25: 953
References 2:
Casoni, F et al J Biol Chem 2005, 280: 16295
References 3:
Han; F et al. Resuscitation 2008; 79: 301
References 4:
Tsuhako H et al. Free radic Biol Med 2010; 48: 704
References 5:
Brunelli L et al. Metabolic brain disease 2012; 27:37
LMR5 reacts with an internal epitope of the LRP/Major Vault Protein (P110), which is strongly overexpressed in various human non-P-glycoprotein MDR tumor cell lines.
Antibody Isotype:
IgG
Monosan Range:
MONOSAN
Clone:
LMR5
Concentration:
250 ug/ ml
Storage buffer:
supernatant with 1% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Scheper RJ et al. Int.J.Cancer 1993; 53: 1475-1479
M2III-6 reacts with an internal epitope of cMOAT/MRP2, a 170-180 kD transmembrane protein known as the canalicular multi-organic anion transporter, absent in patients with the Dubin-Johnson syndrome, an autosomal recessive liver disorder characterized by chronic conjugated hyperbilirubinemia. cMOAT/MRP2 is closely related to the multidrug resistance related protein MRP, and cMOAT/MRP2 overexpression has been observed in a subset of cisplatin resistant cell lines. M2III-6 was raised against a bacterial fusion protein of cMOAB/MRP2, containing the 202-amino acid COOH terminal end of the protein. M2III-6 did not cross react with the human MDR1, MRP1, MRP3 and MRP5 gene products.
M2I-4 reacts with an internal epitope of cMOAT/MRP2, a 170-180 kD transmembrane protein known as the canalicular multi-organic anion transporter, absent in patients with the Dubin-Johnson syndrome, an autosomal recessive liver disorder characterized by chronic conjugated hyperbilirubinemia. cMOAT/MRP2 is closely related to the multidrug resistance related protein MRP, and cMOAT/MRP2 overexpression has been observed in a subset of cisplatin resistant cell lines. M2I-4 was raised against a bacterial fusion protein of cMOAB/MRP2, containing amino acids 215-310 of the protein. M2I-4 did not cross react with the human MDR1, MRP1, MRP3 and MRP5 gene products.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
M2I-4
Concentration:
250 ug/ ml
Storage buffer:
Serum free tissue culture supernatant with 0.7% BSA and 0.1% sodium azide
Mab RNL-1, directed against N-CAM (Neural Cell Adhesion Molecule), reacts with normal neural tissues and endocrine glands, such as pancreatic islet, pituitary gland and adrenal medulla. Expression was also found in Leydig cells of the testis, in the thyroid and in smooth-muscle cells of the small intestine, colon and bladder. The antibody is a valuable marker for the characterization of several neuroendocrine tumors. In immunoblotting (Western) the antibody is reactive with 3 main clusters of protein bands in the molecular weight region of 200 kD, 100 kD, and 25-27 kD. Positive control: Pancreatic islet cells (cell surface staining).
The antibody reacts with an internal epitope of MRP1, a 180-195 kD transmembrane transporter protein overexpressed in various human non-P-glycoprotein MDR tumor cell lines. MRPm5 was raised against a bacterial fusion protein of MRP1, containing amino acids 986-1204 of the protein. MRPm5 does not cross-react with the human MDR1 and MDR3 gene products.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
MRPm5
Concentration:
100 ug/ ml
Format:
Protein G purified
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Cole S et al. Science 1992; 258: 1650-1654
References 2:
Flens M et al. Cancer Res 1994; 54: 4557-4563
References 3:
Zaman et al. Proc Nat Acad Sci 1994; 91: 8822-8826
MRPr1 reacts with an epitope of MRP, a 180-195 kD transmembrane transporter protein overexpressed in various human non-P-glycoprotein MDR tumor cell lines. MRPr1 was raised against a bacterial fusion protein of MRP, containing a segment of 168 amino acids in the amino-proximal half of the protein. MRPr1 does not cross-react with the human MDR1 and MDR3 gene products (Flens et al. 1994).
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
MRPr1
Concentration:
250 ug/ ml
Storage buffer:
Serum free tissue culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Cole S et al. Science 1992; 258: 1650-1654
References 2:
Flens M et al. Cancer Res 1994; 54: 4557-4563
References 3:
Zaman et al. Proc Nat Acad Sci 1994; 91: 8822-8826
MRPm6 reacts with an internal epitope of MRP, a 180-195 kD transmembrane transporter protein overexpressed in various human non-P-glycoprotein MDR tumor cell lines. MRPm6 was raised against a bacterial fusion protein of MRP, containing a segment of 170 amino acids in the carboxy terminal end and part of the carboxy proximal nucleotide binding domain of the protein. MRPm6 does not crossreact with the human MDR1 and MDR3 gene products
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
MRPm6
Concentration:
250 ug/ ml
Storage buffer:
Serum free tissue culture supernatant with 0.7% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Moll I et al. Eur J Cell Biol 2005; 84: 259-271
References 2:
Riedel I et al. Virchows Arch 2001; 438: 181-191
References 3:
Romih R et al. Cell Biol 1998; 109: 263-269
References 4:
Demirkesen C et al. J Cutan Pathol. 1995; 22: 518-535
LRP-56 reacts with an internal epitope of the LRP-protein (P110), which is strongly over-expressed in various human non-P-glycoprotein MDR tumor cell lines.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
LRP-56
Concentration:
100 ug/ml
Storage buffer:
supernatant with 1% BSA and 0.1% sodium azide
Storage:
2-8°C
References 1:
Scheper RJ et al. Int.J.Cancer 1993; 53: 1475-1479
References 2:
List AF et al. Blood 1993; 82: 443a
References 3:
Izquierdo MA et al. ProcAACR 1993; 34: 311
References 4:
Scheper RJ et al. Proc.Am.Ass Cancer Res. 1994; 54: 4557-4563
References 5:
Scheffer GL et al. Proc Am Ass Cancer Res 1995; 36: 323
Acid extracts of boar spermatozoa were subjected to hydrophobic chromatography and the pooled fraction with reactivity to N-alpha benzoylarginine-4-nitroanilide was used for immunization.
Acrosin is a serine proteinase expressed in the acrosome of mature spermatozoa. This enzyme facilitates penetration of the sperm through the zona pellucida of the ovum.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
ACR.2
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The antibody is specific for Synaptophysin. The specificity was ascertained by immunoblotting and immunohistochemistry. The antibody predominantly reacts with a 38 kDa transmembrane glycoprotein from synaptic vesicles.
The antibody recognizes a heterodimeric glycoprotein of 145, 185 kD which has been identified as NCAM (Neural Cell Adhesion Module). Detection of NCAM on cultured cells and in frozen tissue sections. The antibody is further useful for studies on neoplasms of the lung and the nervous system.
The antibody reacts with a conserved cytoplasmic epitope of the plasma membrane-associated 170-180 kD glycoprotein, the expression of which is strongly correlated with the degree of multi-drug-resistance (MDR) derived MDR cell lines and human MDR cell lines, including cell lines derived from lung, ovaries and B cell lymphomas. Target species: Human, cross-reaction: Chinese hamster. No cross-reaction: mouse and rat.
The antibody reacts with a Guinea pig lymphocyte subset probably analog to human CD8 (cytotoxic/suppressor) subset. CD8 comprises 2 subunits, alpha and beta and exists as either an alpha/alpha homodimer or an alpha/beta heterodimer. Sequence suggests that guinea pig CD8 is more closely related to human than rat or mouse CD8. Although this monoclonal originally was developed for the detection of a Guinea pig lymphocyte subset, it also can be used as a negative control for the JSB-1 monoclonal antibodies because it is of the same IgG subclass.
NK cells make up approximately 10% - 25% of peripheral blood lymphocytes. In addition, NCAM is expressed on a variety of neural tissues and some tumors of neuro-endocrine origin, such as small cell lung cancer (SCLC). The CD56 antigen is not expressed on other immune cells. MOC-1 detects an isoform of the neural cell adhesion molecule (NCAM) expressed on natural killer (NK) cells of approximately 145 kDa
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
MOC-1
Concentration:
50 ug/ml
Storage buffer:
0.01 M sodium phosphate, 0.15 M NaCl; pH 7.3, 0.2% BSA, 0.09% sodiumazide
Storage:
2-8°C
References 1:
De Leij, L., et al., 1985. Cancer Research 45: 2192-2200
The antibody is directed against a single band of 180 kD of human carcinoembryonic antigen (CEA) and shows no cross-reactivity neither with bilary glycoprotein (BGP) nor with non-specific cross-reacting antigen (NCA).
The antibody stains over 95% of primary renal cell carcinomas and 60% of metastases of renal cell carcinoma and glomerular visceral epithelium and proximal tubules in normal kidney (cytoplasmic). It does not stain epithelial cells of non-renal malignancies.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
RC38
Concentration:
15 ug/ml
Storage buffer:
Culture medium with 0.01M Sodium Azide
Storage:
2-8°C
References 1:
Oosterwijk E et al. (1986). Am J Pathol 123, 301-309
The antibody recognizes a transmembrane glycoprotein of 140 and 180 kD which has been identified as NCAM (Neural Cell Adhesion Module). At the international Workshop on SCLC antibodies 123C3 has been categorized as cluster 1 antibody. All cells in small cell carcinomas and carcinoids of the lung are strongly positive for 123C3. A minority of cases of other major types of lung carcinoma are sometimes positive as well: however this positivity is generally weak and focal. Adenoid cystic carcinomas of bronchial glands are strongly positive. Neuroblastoma's and Wilms tumors are usually also staining strongly positive. In non-small lung cell carcinomas, 123C3 staining has been associated with more advanced stage and a decreased survival after surgery. Furthermore, this antibody can be used to support diagnosis of lymphoma or to detect residual disease for cases of CD56 positive T/NK -cell lymphoma in which the neoplastic lymphoid cells are small and show minimal atypia, especially in small biopsies.
The antibody stains 80% of non-mucinous primary and metastatic ovarian cancers. This monoclonal antibody is used for the identification of primary and metastatic non-mucinous ovarian carcinoma and ovarian carcinoma cells in peritoneal fluids. It rarely stains nongynaecological malignancies.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
OV632
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.01M Sodium Azide
Storage:
2-8°C
References 1:
Fleuren GJ et al. (1987) Virchows Archiv A, 410, 481-486
References 2:
Boerman OC et al. (1991) Int J Gynecol 10, 15-23
References 3:
Delahye H et al., (1991) J Pathol 165 137-143
References 4:
Boerman OC et al. (1991) Anticancer Res 10, 1289-1296
CD63 (LAMP-3, lysosome-associated membrane protein-3), a glycoprotein of tetraspanin family, is present in late endosomes, lysosomes and secretory vesicles of various cell types. It is also present in the plasma membrane, usually following cell activation. Hence, it has become an widely used basophil activation marker. In mast cells, however, CD63 exposition does not need their activation. CD63 interacts with integrins and affects phagocytosis and cell migration, it is also involved in H/K-ATPase trafficking regulation of ROMK1 channels. CD63 also serves as a T-cell costimulation molecule. Expression of CD63 can be used for predicting the prognosis in earlier stages of carcinomas.
CD63 (LAMP-3, lysosome-associated membrane protein-3), a glycoprotein of tetraspanin family, is present in late endosomes, lysosomes and secretory vesicles of various cell types. It is also present in the plasma membrane, usually following cell activation. Hence, it has become an widely used basophil activation marker. In mast cells, however, CD63 exposition does not need their activation. CD63 interacts with integrins and affects phagocytosis and cell migration, it is also involved in H/K-ATPase trafficking regulation of ROMK1 channels. CD63 also serves as a T-cell costimulation molecule. Expression of CD63 can be used for predicting the prognosis in earlier stages of carcinomas.
CD63 (LAMP-3, lysosome-associated membrane protein-3), a glycoprotein of tetraspanin family, is present in late endosomes, lysosomes and secretory vesicles of various cell types. It is also present in the plasma membrane, usually following cell activation. Hence, it has become an widely used basophil activation marker. In mast cells, however, CD63 exposition does not need their activation. CD63 interacts with integrins and affects phagocytosis and cell migration, it is also involved in H/K-ATPase trafficking regulation of ROMK1 channels. CD63 also serves as a T-cell costimulation molecule. Expression of CD63 can be used for predicting the prognosis in earlier stages of carcinomas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
MEM-259
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Skp2 (S-phase Kinase-associated Protein 2) belongs to the family of F-box proteins that interact with the Cyclin A-Cdk2 complex. Skp2 is essential for the G1-S transition in both transformed cells and diploid fibroblasts. Biochemical and genetic experiments have demonstrated that Skp2 is required for the ubiquitination and consequent degradation of p27 in cultured mammalian cells and in vitro reconstitution assays. In normal tissues, Skp2 is expressed in tonsil and placenta.
Mouse anti Human macrophages, clone MAC387 recognizes the L1 or Calprotectin molecule, an intracytoplasmic antigen comprised of a 12 kDa alpha chain and a 14 kDa beta chain. Although originally described as binding to epitopes common to both the alpha and beta chains (Flavell et al. 1987) subsequent studies indicate that the antibody detects an epitope exclusively expressed on the beta chain (Goebeler et al. 1994) demonstrated by immunofluorescent and western blotting on both naturally expressing and transfected targets. In addition Mouse anti Human macrophages, clone MAC387 detects the beta chain in complex with the alpha.The antigen recognized by Mouse anti Human macrophages, clone MAC387 is expressed by granulocytes, monocytes and by tissue macrophages. Variable results have been reported for staining brain macrophages and microglia. The epitope recognized appears to be well conserved and the antibody is routinely used for the detection of myeloid cells in a wide range of species.
Mouse anti human CD236 antibody, clone HEA-125 recognizes CD326 also known as Adenocarcinoma-associated antigen, Epithelial glycoprotein 314 or Epithelial Cell Adhesion Molecule (Ep-CAM). CD326 is a 314 amino acid ~34 kDa single pass type I transmembrane glycoprotein nearing a single thyroglobulin type-1 domain (UniProt: P16422).CD326 is expressed on the basolateral membrane of cells by the majority of epithelial tissues, with the exception of adult squamous epithelium and some specific epithelial cell types including hepatocytes and gastric epithelial cells.CD326 expression has been reported to be a possible marker of early malignancy, with expression being increased in tumour cells (Balzar et al. 1999), and de novo expression being seen in dysplastic squamous epithelium (Winter et al. 2003). CD326 has been identified independently by a number of groups, and it has been known by a variety of names including Epithelial Specific Antigen, MOC31 (Proca et al. 2000) and Ber-EP4 (Patriarca et al. 2012).
Mouse anti Human TGF beta antibody, clone TB21 recognizes both human platelet-derived and recombinant TGF-beta1 in enzyme-linked immunosorbent assay (ELISA). Mouse anti Human TGF beta antibody, clone TB21 demonstrates neutralising activity against TGF-beta1 in cell proliferation assays. Mouse anti Human TGF beta antibody, clone TB21 has been demonstrated to react with dimeric (~25 kDa) or monomeric (~12.5 kDa) molecules of natural TGF-beta1 under non-reducing and reducing conditions respectively.
Mouse anti Human Prolactin antibody, clone INN-hPRL-1 recognises human prolactin, also known as luteotropic hormone or luteotropin, binding to epitope 'c' as determined by a competitive binding assay (Staindl et al. 1987). Prolactin is a 199 amnio acid ~24 kDa secreted anterior pituitary hormone acting to promote lactation.
Mouse anti HumanFSH beta 2, clone INN-hFSH-60 is directed against the Beta 2 epitope of the Beta subunit of hFSH, INN-hFSH-60 shows strong reactions with hFSH and beta-hFSH, but no cross-reactivity with hTSH, hLH, hCG, alpha-hCG or alpha-hFSH. The recognition site is located near the alpha 1 binding site on the hFSH molecule. It is not compatible with other anti-hFSH-beta antibodies.
Mouse anti Human granzyme A antibody, clone GA6 recognizes Granzyme A, a ~60 kDa disulphide-linked homodimeric protein of two 262 amino acid chains, expressed in cytoplasmic granules of cytotoxic lymphocytes and NK cells.Granzyme A is involved in the induction of apoptosis via its activity as a serine protease, but this would seem to be subsidiary to the role of Granzyme B. Granzyme A deficient mice are indistinguishable from normal animals in their response to infection.Granzyme A has been proposed as a potential biomarker for patients with active tuberculosis with significantly lower levels present in the plasma of patients with the active form of the disease compared to patients with latent infection (Guggino et al. 2015).
Mouse anti Human aquaporin 1 antibody, clone 1/A5F6 recognizes an epitope within the cytoplasmic domain of the water-specific channel aquaporin 1, also known as AQP1 or CHIP-28.Aquaporin 1 is a ~28 kDa integral membrane protein which was originally identified in red blood cells and the kidney. AQP1 is also expressed by the choroid plexus and various other tissues. The glycosylated forms of AQP1 range between 40-60 kDa.
Among the six actin isoforms described in mammals, two are found in virtually all cells (?- and ?-cytoplasmic), two are detected in smooth muscle cells (?- and ?-smooth muscle) and two are present in striated muscles, one predominantly in skeletal (?-skeletal) and one in cardiac (?-cardiac) muscle cells. These actin isoforms differ slightly in their N-terminus, but the sequence of each of these actins is highly conserved in higher vertebRates. Alpha- muscle actin is present in striated as well as smooth muscle cells, and in pathological tissues derived therefrom.HHF35 reacts with both ?-muscle and ?-smooth muscle actin, and therefore reacts with skeletal muscle, cardiac muscle, vascular and visceral smooth muscle cells, pericytes and myoepithelial cells. It is also reactive in myofibroblasts. It does not react with epithelial, endothelial, neural or normal connective tissue cells when applied under the proper conditions to these tissue sections.
Mouse anti Human Mcm5 antibody, clone CRCT5.1 recognizes human Mcm-5 (minichromosome maintenance protein 5), also known as DNA replication licensing factor MCM5 or P1-CDC46. Mcm5 is a nuclear protein of ~95kDa with an important role in the control of DNA replication (Snyder et al. 2005).Immunocytochemical assessment of Mcm5 expression may be of value in improving the accuracy of cervical smear testing for the detection of malignancy (Murphy et al. 2004).
9-13M1 recognizes the peptide core of gastric mucin M1/MUC5AC), and more specifically with the d epitope amongst the a, b, c, d, e, f, g and h protein core epitopes defined by Bara for M1. 9-13M1 and 2-11M1 react exclusively with epitopes located in the the Nterminal cysteine-rich part of the peptide core MUC5AC. MUC5AC is present in primary ovarian mucinous cancer and gastric cancer, but usually absent in colorectal adenocarcinoma, thus showing an expression pattern opposite to MUC2. Anti-MUC5AC may be useful for differential identification of primary mucinous ovarian tumors from colon adenocarcinoma metastatic to the ovary. MUC5AC antibodies may also be useful for identification pancreatic carcinoma and pre-cancerous changes vs. normal pancreas
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
9-13M1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al., Cancer Res.46: 3983-3989 (1986)
References 2:
Bara, J. et al., Biochem. J. 254: 185-193 (1988)
References 3:
Bara, J. et al., Int. J. Cancer 47: 304-310 (1991)
References 4:
Bara, J. et al., J. Immunol. Methods 149: 105-113 (1992)
References 5:
Guyonnet Duperat V. et al., Biochem. J. 305: 211 219 (1995)
58M1 recognizes the peptide core of gastric mucin M1 (now: MUC5AC), and more specifically with the e epitope amongst the a, b, c, d, e, f, g and h protein core epitopes defined by Bara for M1. MUC5AC is present in primary ovarian mucinous cancer and gastric cancer, but usually absent in colorectal adenocarcinoma, thus showing an expression pattern opposite to MUC2. Anti-MUC5AC may be useful for differential identification of primary mucinous ovarian tumors from colon adenocarcinoma metastatic to the ovary. MUC5AC antibodies may also be useful for identification pancreatic carcinoma and pre-cancerous changes vs. normal pancreas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
58M1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al., Cancer Res.46: 3983-3989 (1986)
References 2:
Bara, J. et al., Biochem. J. 254: 185-193 (1988)
References 3:
Bara, J. et al., Int. J. Cancer 47: 304-310 (1991)
References 4:
Bara, J. et al., J. Immunol. Methods 149: 105-113 (1992)
References 5:
Guyonnet Duperat V. et al., Biochem. J. 305: 211 219 (1995)
45M1 recognizes the peptide core of gastric mucin M1 (now: MUC5AC), and more specifically with the h epitope amongst the a, b, c, d, e, f, g and h protein core epitopes defined by Bara for M1. 45M1 and 2-12M1 both specifically react with epitopes located in the C-terminal cysteine rich part of the peptide core of MUC5AC. MUC5AC is present in primary ovarian mucinous cancer and gastric cancer, but usually absent in colorectal adenocarcinoma, thus showing an expression pattern opposite to MUC2. Anti-MUC5AC may be useful for differential identification of primary mucinous ovarian tumors from colon adenocarcinoma metastatic to the ovary. MUC5AC antibodies may also be useful for identification pancreatic carcinoma and pre-cancerous changes vs. normal pancreas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
45M1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al., Int. J. Cancer 47: 304-310 (1991)
References 2:
Bara, J. et al., J. Immunol. Methods 149: 105-113 (1992)
2-12M1 recognizes the peptide core of gastric mucin M1/MUC5AC, and more specifically with the c epitope amongst the a, b, c, d, e, f, g, and h protein core epitopes defined by Bara for M1. 2-12M1 and 45M1 both specifically react with epitopes located in the Cterminal cysteine rich part of the peptide core of gastric mucin (MUC5AC). MUC5AC is present in primary ovarian mucinous cancer and gastric cancer, but usually absent in colorectal adenocarcinoma, thus showing an expression pattern opposite to MUC2. AntiMUC5AC may be useful for differential identification of primary mucinous ovarian tumors from colon adenocarcinoma metastatic to the ovary. MUC5AC antibodies may also be useful for identification pancreatic carcinoma and pre-cancerous changes vs. normal pancreas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
2-12M1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al., Cancer Res.46: 3983-3989 (1986)
References 2:
Bara, J. et al., Biochem. J. 254: 185-193 (1988)
References 3:
Bara, J. et al., Int. J. Cancer 47: 304-310 (1991)
References 4:
Bara, J. et al., J. Immunol. Methods 149: 105-113 (1992)
References 5:
Guyonnet Duperat V. et al., Biochem. J. 305: 211 219 (1995)
2-11M1 recognizes the peptide core of gastric mucin M1/MUC5AC, and more specifically with the b epitope amongst the a, b, c, d, e, f, g, and h protein core epitopes defined by Bara for M1. 2-11M1 and 9-13M1 react exclusively with epitopes located in the N-terminal cysteine-rich part of the peptide core MUC5AC. MUC5AC is present in primary ovarian mucinous cancer and gastric cancer, but usually absent in colorectal adenocarcinoma, thus showing an expression pattern opposite to MUC2. Anti-MUC5AC may be useful for differential identification of primary mucinous ovarian tumors from colon adenocarcinoma metastatic to the ovary. MUC5AC antibodies may also be useful for identification pancreatic carcinoma and pre-cancerous changes vs. normal pancreas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
2-11M1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al., Cancer Res.46: 3983-3989 (1986)
References 2:
Bara, J. et al., Biochem. J. 254: 185-193 (1988)
References 3:
Bara, J. et al., Int. J. Cancer 47: 304-310 (1991)
References 4:
Bara, J. et al., J. Immunol. Methods 149: 105-113 (1992)
References 5:
Guyonnet Duperat V. et al., Biochem. J. 305: 211 219 (1995)
1-13M1 recognizes the peptide core of gastric mucin M1/MUC5AC), and more specifically with the a epitope, which is the most abundant amongst the a, b, c, d, e, f, and h protein core epitopes defined by Bara for M1. MUC5AC is present in primary ovarian mucinous cancer and gastric cancer, but usually absent in colorectal adenocarcinoma, thus showing an expression pattern opposite to MUC2. Anti-MUC5AC may be useful for differential identification of primary mucinous ovarian tumors from colon adenocarcinoma metastatic to the ovary. MUC5AC antibodies may also be useful for identification pancreatic carcinoma and precancerous changes vs. normal pancreas.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
1-13M1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Bara, J. et al., Cancer Res.46: 3983-3989 (1986)
References 2:
Bara, J. et al., Biochem. J. 254: 185-193 (1988)
References 3:
Bara, J. et al., Int. J. Cancer 47: 304-310 (1991)
References 4:
Bara, J. et al., J. Immunol. Methods 149: 105-113 (1992)
References 5:
Guyonnet Duperat V. et al., Biochem. J. 305: 211 219 (1995)
EBS-C-002 reacts with NuMA or Nuclear Mitotic Apparatus protein, which at the onset of mitosis redistributes from the nucleus to two centrosomal structures at the poles of the mitotic spindle, where it plays a vital role in establishing and maintaining its bipolar structure. After anaphase the protein redistributes from the spindle polar region into the reforming nucleus and concentrates initially at the site where nuclear lamins and perichomatin have been reported to assemble. In contrast to mitotic cells, post-mitotic neurons display NuMA both in the nucleus and in the cytoplasm. Due to release from dead cells, NuMA is also used as oncological marker in serum and urine. In addition, chromosomal translocation of this gene with the RARA (retinoic acid receptor, alpha) gene on chromosome 17 has been detected in patients with acute promyelocytic leukemia.
VU-2G7 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU2G7 includes the PDTR motif, located in the VNTR domain of MUC1. Binding of VU-2G7 is significantly enhanced when the threonine of the PDTR motif bears a GalNAc
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
2G7
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Ryuko, K. et al. Tumor Biol. 21(4): 197-210 (2000)
References 2:
Karsten, U. et al. Cancer. Res. 58(12): 2541-2549 (1998)
VU-13F11 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU-13F11 has not yet been determined but is located in the VNTR domain of MUC1 (confirmed by ELISA).
Antibody Isotype:
IgM
Monosan Range:
MONOSAN
Clone:
VU-13F11
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Schol D. et al, Tumor Biol. 19(Suppl 1): 35-45 (1998)
VU-12E1 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU-12- E1 is PDTRPAP, located in the VNTR domain of MUC1. Binding of VU-12E1 is enhanced after glycosylation of the DTR motif
VU-11E2 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU11E2 is TSAPDTRP, located in the VNTR domain of MUC1. Binding of VU-11E2 is enhanced after glycosylation of the DTR motif
VU-11D1 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU11D1 is TSAPDTRP, located in the VNTR domain of MUC1. Binding of VU-11D1 is enhanced after glycosylation of the DTR motif.
VU-4H5 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU-4H5 is PDTR, located in the VNTR domain of MUC1. In tissue sections, VU-4H5 also displays prominent staining of the cytoplasm.
VU-3D1 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU3D1 is SAPDTRPA, located in the VNTR domain of MUC1.
VU-3C6 reacts with the protein core of MUC1, an apical cell side epithelial marker which is upregulated or switched on in the majority of carcinomas. The dominant epitope of VU-3- C6 is GVTSAPDTRPAP, located in the VNTR domain of MUC1. Binding of VU-3C6 is enhanced after glycosylation of the DTR motif.
This monoclonal antibody binds to human MMP-2. Reactivity in other species has not been determined, but the 13-mer peptide used for the immunization shows a 100% match with rabbit MMP-2, and differs on only 1 amino acid with rat and mouse MMP-2. The antibody was tested for cross-reactivity with MMP-1, MMP-3, and MMP-9 and showed mild cross-reactivity with MMP-3.
This monoclonal antibody binds to human MMP-1. Reactivity in other species has not been determined. The antibody was tested for cross-reactivity with MMP-2, MMP-3, and MMP-9 and did not cross-react.
NHERF1 (Na+/H+ exchanger regulatory factor 1), also known as EBP50 (ezrin, radixin, moesin-binding phosphoprotein 50) is an adaptor protein, which associates with beta-catenin and is required for its localization at the cell-cell junctions, interacts with various G protein-coupled receptors and regulates their traffic, as well as sodium-hydrogen exchange and sodium-dependent phosphate transport. NHERF1/EBP50 inhibits cell motility and is required to suppress anchorage-independent growth. It contains C-terminal ERM (ezrin, radixin, moesin)-binding region and two N-terminal PDZ (postsynaptic-density-95/disc-large/ZO1 homology) domains and is able to form head-to-tail intramolecular conformation to regulate its interactions.
Antibody Isotype:
IgG1 kappa
Monosan Range:
MONOSAN
Clone:
EBP-10
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Monoclonal Anti-IDH1 (R132H) recognizes only the R132H mutation of human IDH1 (R132H) and does not cross react with other mutations. The most frequent known mutation (>90%) is the alteration of arginine to histidine (R132H).6. Hence, antibodies that recognize the IDH1R132H mutation can be useful for the diagnosis of mutation-bearing tumors like gliomas. Positive control Human Glioma tissue
Antibody Isotype:
IgG1k
Monosan Range:
MONOSAN
Clone:
Hmab-1
Concentration:
lot specific
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Capper et al. Pathol 2010;20(1):245-254
References 2:
Capper et al. Am J Surg Pathol 2010;34(8):1199-1204
P16 is a mitotic inhibitor protein. It competes with D-type cyclins to bind to cdk4 and cdk6. It acts as tumor suppressor and inhibits the progression of cells through the G1 phase of the cell cycle. Positive Control Tissue Uterine cervical squamous cell carcinoma
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
JC2
Concentration:
lot specific
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Sherr et al. Cold Spring Harb Symp Quant Biol 1994;59:11-19
PTEN gene is a tumor suppressor gene that maps to chromosome 10q23. PTEN, a novel tumor suppressor, functions as a regulator of both cell cycle progression and apoptosis ). Potentially, mutation and deletion of PTEN gene may result in a new signal transduction pathway related to human malignant tumors. Studies have demonstrated a reduction of PTEN expression in advanced breast cancers. Positive control tissue Breast, Renal Cell and Prostate carcinomas
The programmed death receptor 1 (PD-1) protein is a cell-surface receptor on certain lymphocytes that, with its ligand programmed death ligand 1 (PD-L1), helps to down-regulate immune responses. Many cancer types express PD-L1 and evade immune recognition via the PD-1/PD-L1 interaction. Precision therapies targeting the PD-1/PD-L1 pathway have the potential to improve response and thereby offer a novel treatment avenue to some patients with cancer.
INSM1 (insulinoma-associated protein 1), also known as zinc-finger protein IA-1, is a developmentally regulated zinc-finger transcription factor. It localizes to the nucleus and is expressed in embryonic tissues undergoing neuroendocrine differentiation. INSM1 is not expressed in normal adult tissues but it can be found highly expressed in neuroendocrine tumors. INSM1 contains five Cys2-His2-type zinc-finger DNA binding domains and a prohormone domain. INSM1 acts as a transcriptional repressor of the Neuro D promoter and recruits cyclin D1 as a corepressor. It plays an important role in neuroendocrine development and is required for normal differentiation of pancreatic endocrine cells. Inhibition of INSM1 results in decreased formation of glucagon and Insulin positive cells. The gene encoding INSM1 is directly regulated by Neurogenin 3 which binds chromatin in the INSM1 promoter region and induces transcription.
Antibody Isotype:
IgG1k
Monosan Range:
MONOSAN
Clone:
A-8
Concentration:
lot specific
Storage buffer:
Tissue culture supernatant with sodium azide
Storage:
2-8°C
References 1:
Li et al. Biochem Biophys Res Comm 1997;236;776-781
References 2:
Breslin et al. Nucleic Acids Res 2002;30:1038-1045
Monoclonal antibody aE11 reacts with a C9 neoantigen of the terminal complement complex (TCC). The three distinct activation pathways of complement converge with the formation of a C5 convertase. The cleavage of C5 by this convertase initiates the lytic or terminal pathway. In contrast to the activation pathways, which require enzymatic cleavage for activation, the terminal pathway relies on conformational changes induced by binding. Binding of C6 facilitates binding of C7 which alters the conformation of the complex. After binding of C8, a variable number of C9 molecules associate with the C5b678 complex, which is also termed- the terminal complement complex (TCC). The formation of TCC causes lysis of cells or can trigger a variety of cellular metabolic pathways resulting in the synthesis and release of inflammatory mediators. The TCC contains neoantigens that are absent from the individual native components.- C9 neoantigens are present both in the membrane-bound (MAC) and the fluid-phase (SC5b-9) complex. TCC is present in normal human plasma and increased in patients with complement activation.
EBS-CD-044 reacts with CD99 or MIC2. Human thymocytes, PBLs and some T-ALL isolates and cell lines are positive. It is also present on pancreas and on Ewing sarcoma, which forms the practical application of the antibody. It is involved in T-cell adhesion processes and in spontaneous rosette formation with erythrocytes.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-044
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Rajasekariah et al. in: Leukocyte Typing III A. McMichael (ed), Oxford University Press, Oxford (1987)
Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in Human epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 7.8. The individual Human Cytokeratins are numbered 1 to 20. The various epithelia in the Human body usually express Cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used in the distinction of different types of epithelial malignancies. The Cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays. RCK105 reacts exclusively with Cytokeratin 7 which is present in a subgroup of glandular epithelia and their tumors, as well as transitional epithelium and transitional carcinoma.
EBS-T-232 reacts with SITTTE in the VNTR domain of human MUC3. The mucins are a family of highly glycosylated, secreted proteins with a basic structure consisting of a variable number of tandem repeats (VNTRs) encoded by 60 base pairs (MUC1), 69 base pairs (MC2) and 51 base pairs (MUC3). Cancer cells of colon, breast and stomach, normal cells of salivary gland, breast, lung, and gastrointestinal tract are positive for MUC3.
Antibody Isotype:
IgG2b-K
Monosan Range:
MONOSAN
Clone:
EBS-T-232
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Apostolopoulos, V. et al. J. Gastroenterol. Hepatol. 1995; 10 (5): 555-561
CD36 is a 80-90 kDa protein, expressed on platelets, monocytes and macrophages, microvascular endothelial cells, erythrocyte precursors, mammary epithelial cells, and some macrophage derived dendritic cells. CD36 acts as a receptor for thrombospondin (TSP), collagen types I, IV and V, P.falciparum malaria-infected erythrocytes, and sickle erythrocytes. CD36 plays a role in platelet aggregation, macrophage foam cell development, inflammation, and the tissue ischemia observed in sickle cell disease and cerebral malaria.
203-6 Reacts with human CD27, a disulphide-linked 120 kDa dimer. CD27 is a lymphocyte-specific member of the TNF-R/NGF-R superfamily, and is expressed on a subset of human thymocytes and on the majority of mature T-lymphocytes. CD27 is highly expressed on activated T and B-cells. 203- 6 was clustered at the VIth WLDA.
Antibody Isotype:
IgG3-K
Monosan Range:
MONOSAN
Clone:
203-6
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kishimoto T. et al., eds. Leukocyte Typing VI, p509-514, Garland Publishing, Inc, (1997)
Ep-CAM (also called ESA, EGP40, 17-1A antigen, KSA, GA7333-2) is a 40 kDa epithelial protein expressed on baso-lateral cell surfaces in very many epithelial tissues (but absent from mesothelial tissues). The 324AA have 3 potential glycosylation sites and is a transmembrane glycoprotein. The extracellular domain has a cysteine-rich repeat and a small domain with homology to nidogen. It is a homophilic cell-cell adhesion molecule (Ep-CAM). EBS-CD-061 reacts with most epithelial cells and carcinomas.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-061
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Edwards DP et. al. Cancer Res 46:1306-17 (1986)
References 2:
Litvinov et al. J. Cell. Biol. 125: 437-446 (1994)
EBS-CD-060 reacts with a peptide epitope on the extracellular domain of human Glycophorin A, a 39 kDa sialoglycoprotein, present on red cells and erythroid precursor cells. Glycophorin A is the carrier of blood group M and N specificities, while Glycophorin B carries S and U specificities. Providing a mucin like coat, Glycophorin may play a role in preventing red cell aggregation in the circulation. Glycophorin also acts as receptors for Sendai and Parvovirus
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-060
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Cartron JP et al, Transfus Med Rev 6(2): 63-92 (1992)
CD109 is a GPI-anchored member of the alpha-2-macroglobulin (A2M) and complement family of proteins. It is expressed on activated T-cells, platelets, hematopoietic stem cells, megakaryocyte precursors, vascular endothelial cells, basal and myoepithelial cells of secretory glands, and squamous cell carcinomas. A 170-180 kDa precursor is autocatalytically reduced to 150 kDa and 120 kDa forms. On keratinocytes CD109 binds TGF-beta and associates with TGFbeta RI and TGF-beta RII, resulting in inhibition of TGF-beta signalling. Polymorphisms of CD109 include the platelet-specific Gov antigen and the blood group ABH antigens. Alloantibodies directed against these antigens result in unsuccessful platelet transfusions, neonatal alloimmune thrombocytopenia, and post-transfusion purpura.
EBS-CD-045 reacts with human CD100, a 150 kDa homodimer cell-surface antigen that is expressed on resting and PHA-stimulated T-cells. It is absent from bone marrow, erythrocytes, eosinophils and endothelial cells. The protein is weakly expressed on NK-cells, EBV transformed B-cells, monocytes and tumor T-cell lines. It plays a role in homotypic cell adhesion and in T-cell activation.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-045
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Hall K, et al. P. Natl. Acad. Sci. USA 93: 11780 (1996)
References 2:
Mizrahi S, et al. PLoS One. 2(9): e818 (2007).
References 3:
Yoshino N, et al.. Exp. Anim. (Tokyo) 49: 97 (2000)
EBS-CD-007 recognized a protein of 32 kDa, identified as CD8b. The CD8 molecule consists of ? and ? chains, which are disulphide-linked into heterodimers or homodimers. CD8 is expressed on a T-cell subset (cytotoxic-suppressor T-cells), thymocytes and NK cells. The majority of CD8+ T-cells express CD8 as heterodimer. Some subpopulation of CD8+ T-cells as well as NKcells may express homodimer. CD8 functions as a co-receptor is concert with TCR for binding the MHC class I/peptide complex. HIV-2 envelope glycoprotein binds CD8 ? chain but not ? chain.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-007
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. et. al. Leukocyte Typing IV, p342-343, Oxford University Press, 1989
References 2:
Parnes JR, Semin Immunol 6: 221-229 (1994)
References 3:
Hadi Hossein Nataj Arab et al., Gastroenterol Hepatol Bed Bench. 8(2): 132139 (2015)
IBL-3/25 is directed against the ?-chain of CD8. The CD8 complex consists of a disulphide-linked ?/? heterodimer with a MW of 30 kDa or an ?/? homodimer with a MW of 32 kDa. The CD8 molecule binds to HLA class I molecules during interaction of CD8+ T-cells with antigenpresenting cells or with target cells. CD8+ T-cells include most of the cytotoxic T-cells.
Antibody Isotype:
IgG-K
Monosan Range:
MONOSAN
Clone:
IBL-3/25
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. et. al. Leukocyte Typing IV, p342-343, (1989)
References 2:
Parnes JR, Semin Immunol 6: 221-229 (1994)
References 3:
Delon J. et al. Immunity 9(4): 467-73 (1998)
References 4:
Akimoto H, et al. Immunology 95(2): 214-218 (1998)
143-44 recognizes a protein of 32 kDa, identified as CD8a. The CD8 molecule consists of ? and ?-chains, which are disulphide-linked into heterodimers or homodimers. CD8 is expressed on a T-cell subset (cytotoxic/suppressor T-cells), thymocytes and NK-cells. The majority of CD8+ T-cells express CD8 as heterodimer. Some subpopulation of CD8+ T-cells as well as NK-cells may express homodimer. CD8 functions as a co-receptor in concert with TCR for binding the MHC class I/peptide complex. The HIV-2 envelope glycoprotein binds CD8 ?-chain but not ?-chain. 124-1D1 was assigned at the IVth International Workshop.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
143-44
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp, W. et. al. Leucocyte Typing IV, p1076, Oxford Univ. Press (1989)
EBS-CD-009 recognizes a protein of 32 kDa, identified as CD8a. The CD8 molecule consists of ? and ?chain, which are disulphide-linked into heterodimers or homodimers. CD8 is expressed on a T-cell subset (cytotoxic/suppressor T-cells), thymocytes and NK-cells. The majority of CD8+ T-cells express CD8 as heterodimer. Some subpopulation of SD8+ T-cells as well as NK-cells may express homodimer. CD8 functions as a co-receptor in concert with TCR for binding the MHC class I/peptide complex. HIV-2 envelope glycoprotein binds CD8 ?-chain but not ?-chain.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-009
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. et. al. Leukocyte Typing IV, p342-343, (1989)
References 2:
Parnes JR, Semin Immunol 6: 221-229 (1994)
References 3:
Delon J. et al. Immunity 9(4): 467-73 (1998)
References 4:
Akimoto H, et al. Immunology 95(2): 214-218 (1998)
RIV11 recognizes a protein of 32 kDa, identified as CD8a. The CD8 molecule consists of ? and ? chains, which are disulphide-linked into heterodimers or homodimers. CD8 is expressed on a T-cell subset (cytotoxic/suppressor T-cells), thymocytes and NK-cells. The majority of CD8+ T-cells express CD8 as heterodimer. Some subpopulations of CD8+ T-cells as well NK-cells may express homodimer. CD8 functions as a co-receptor in concert with TCR for binding the MHC class I/peptide complex. HIV-2 envelope glycoprotein binds CD8 ? chain but not CD8 ? chain.
EBS-CD-42 reacts with CD86, a member of the immunoglobulin superfamily, and highly expressed on monocytes, dendritic cells and stimulated B-cells. It is probably the major CD28 ligand. Furthermore, EBS-CD-042 blocks binding of soluble CD152 to CD86.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-042
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Giguère JF et al, J Virol 78: 6222-32 (2004)
References 2:
Mauri D et al, J Immunol 155: 118-27 (1995)
References 3:
Schlossman S. et al. (eds) Leukocyte Typing V, Oxford University Press (1995)
References 4:
Kishimoto T. et al., eds. Leukocyte Typing VI, p509-514, Garland Publishing, Inc, (1997)
References 5:
Sandilands GP et al, Immunology 108: 329337 (2003)
CD80 is involved in the costimulatory signal essential for T-lymphocyte activation. T-cell proliferation and cytokine production is included by the binding of CD28, binding to CTLA-4 has opposite effects and inhibits T-cell activation.
Antibody Isotype:
IgG2b-K
Monosan Range:
MONOSAN
Clone:
C80-99
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Peach, R.J., et al. J. Biol. Chem. 270: 21181-21187 (1995)
References 2:
Fargeas, C.A., et al. J. Exp. Med. 182: 667-675 (1995)
The transferrin receptor is a type II membrane glycoprotein existing as a homodimer of 180- 190 kDa with interchain disculphide bonding. The ligand is the serum iron transport protein transferrin. CD71 is expressed weakly on all resting leucocytes but is upregulated on all cells upon activation, reflecting the iron dependence of proliferation. In other tissues CD71 is expressed on most dividing cells, but also strongly on brain endothelium and alveolar macrophage. CD71 expression can reflect clinical behaviour or response to therapy in a number of malignancies including leukemia, lymphoma and breast cancer.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-040
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Van de Rijn et al. Cytogenet.Cell Genet. 36: 525 (1983)
References 2:
Oudermans et al. Cancer. 58: 1252 (1986)
References 3:
Beguin Y, et al, Leukemia (12): 2019-25 (1993)
References 4:
Rittenhouse-Diakun K, et al. Photochem Photobiol. 61(5): 523-8 (1995)
124-1D1 recognizes 40 kDa CD7, a member of the immunoglobulin gene superfamily and expressed on the majority of immature and mature T-lymphocytes, and T-cell leukemia. It is also found on natural killer cells, a small subpopulation of normal B-cells and on malignant B-cells. CD7 associates directly with phospoinositol and tyrosine phosphorylation. 124-1D1 was assigned at the IVth International Workshop.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
124-1D1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp, W. et. al. Leucocyte Typing IV, p541, 667-670, 1087, Oxford Univ. Press (1989)
BF12 recognizes 40 kDa CD7, a member of the immunoglobulin gene superfamily and expressed on the majority of immature and mature T-lymphocytes, and T-cell leukemia. It is also found on natural killer cells, a small subpopulation of normal B-cells and on malignant B-cells. CD7 associates directly with phosphoinositol 3-kinase. CD7 ligation induces production of D-3 phosphoinositides and tyrosine phosphorylation.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
BF12
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Wang, MY et. al, Bone Marrow Transplant. 9(5): 319-23 (1992)
CB-30 reacts with CD66e or CEA with MW of 80-200 kDa. CEA is present in fetal gut and is re-expressed in increased amounts in intestinal carcinomas and several other tumors. CEA is not found in benign glands, stroma, or malignant prostatic cells. Antibody to CEA is useful in detecting early foci of gastric carcinoma and in distinguishing pulmonary adenocarcinomas (60-70% are CEA+) from pleural mesotheliomas (rarely or weakly CEA+). Ant-CEA positivity is seen in adenocarcinomas from the lung, colon, stomach, esophagus, pancreas, gallbladder, urachus, salivary gland, ovary, and endocervix.
EBS-CD-036 reacts with CD63 and is mainly used in combination with EBS-CD-147 and/or anti-PEM (MUC1) to identify melanoma from carcinoma in paraffin sections. Melanomas are EBS-CD-036 and EBS-CD-147 positive, but PEM negative. EBS-CD-036 reacts in frozen sections with melanoma and breast cancers, smooth muscle and lung (weakly). In paraffin sections melanomas (primary skin, uveal and choroidal), melanoma metastases, clear cell CA, carcinoids, skin nevi, medullary CA of thyroid, prostate CA, some breast, ovary, lung, colorectal and bladder CA positive. Normal tissues that are positive include: mast cells, sweat glands, Islets of Langerhans, pituitary, pancreas, peribronchial glads, Paneth cells and prostate glands.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-036
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Vennegoor C. et al., Int. J. Cancer 35: 287-295 (1985)
References 2:
Palmer AA et al., Pathology 17: 335-339 (1985)
References 3:
Hagen EC et al., Histopathology 10: 689-700 (1986)
References 4:
Duffield, A., et al. Proc. Natl. Acad. Sci. USA 100: 15560-15565 (2003)
EBS-CD-034 reacts with a complex of CD41 and CD61, i.e. alpha IIb integrin and the integrin beta chain. The MW of the GPIIIa is 105 kDa unreduced and 90 kDa reduced. The integrin beta 3 chain can also form a complex called the vitronectin receptor with integrin alpha V: the CD51/CD61 complex. Ligands are fibrinogen, fibronectin, von Willebrand factor, vitronectin and thrombospondin. Residues 237-248 of GPIIIa or CD61 are critical in adhesive protein binding. The CD51/CD61 complex is also found on endothelial cells, some B-cells, monocytes/macrophages and tumor cells.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-034
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Burns et al., Cell 45, 269 280 (1986)
References 2:
McMichael AJ et al. (eds) Leukocyte Typing III, Oxford University Press, Oxford, (1987)
References 3:
Schlossman S. et al. (eds) Leukocyte Typing V, Oxford University Press (1995)
EBS-CD-005 recognizes the CD6 molecule, a single chain transmembrane glycoprotein of 120 kDa, which is expressed on the majority of mature T-cells an mature thymocytes. A B-cell subset is weakly positive and B-CLLs may also be reactive. CD6 is a type 1 transmembrane glycoprotein that is tyrosine phosphorylated during TCR-mediated T-cell activation. CD6 shows significant homology to CD5. Antibodies to CD6 are used to deplete T-cells from bone marrow transplants to prevent graft versus host disease.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-005
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Yssel et al., Cell. Immunol. 105: 161 (1987)
References 2:
Kamoun et al., J. Immunol. 127: 987 (1981)
References 3:
Reinherz et al., Proc. Nat. Acad. Sci. 79: 6047 (1982)
CD59, or protectin, is a 18-22 kDa cell surface molecule on an GPI anchor. It regulates complementmediated cell lysis and is supposed to protect normal and tumor cells from cytotoxic attack by homologous complement through binding to C8 and C9. CD59 is expressed on leucocytes, vascular epithelium, a variety of epithelial cells and placenta. B-cell express low levels. The expression of CD59 on erythrocytes is important for their survival. Genetic defects in GPI-anchor attachment, that cause a reduction or loss of CD59 and CD55 on erythrocytes produce the symptoms of the disease Paroxysmal nocturnal hemoglobinuria (PNH). 193-27 Was typed at the VIth International Workshop on human leucocyte differentiation antigens.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
193-27
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kishimoto T. et al., eds. Leukocyte Typing VI, p509-514, Garland Publishing, Inc, (1997)
References 2:
Shichishima T. et al. Br J Haematol, 85(2):378-386 (1993)
References 3:
Navenot JM. et al. Transfusion 38(4):337-342 (1998)
References 4:
Murray EW et al, J Biol Chem, 273(39):25279-25284 (1998)
EBS-CD-033 reacts with an extracellular domain (close to the cell membrane) of CD56/NCAM. Only 2 isoforms of NCAM are recognized by EBS-CD-033 (180 kDa and 145 kDa). EBS-CD-033 was used successfully for immunoscintigraphy and immunotherapy of SCLC xenografts in nude mice. EBS-CD-033 recognizes NCAM in retinoblastoma, medulloblastomas, astrocytomas, neuroblastomas, and small cell lung carcinomas. NCAM is also expressed on some mesodermally derived tumors (a.o. rhabdomyosarcoma). Anti-CD56 plays an important role in the diagnosis of nodal and nasal NK/T-cell lymphomas.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-033
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Schol DJ. et al, Int. J. Cancer Suppl.2: 35-40 (1988)
References 2:
Kibbelaar RE. et al, Eur J Cancer 27(4): 431-5 (1991)
References 3:
Brezicka FT. et al, APMIS. 99(9): 797-802 (1991)
References 4:
Takaku H. et al. Am J Gastroenterol. 94(5): 1402-4 (1999)
References 5:
Kaufman O. et al, Hum. Pathol. 28(12): 1373-1378 (1997)
F4-29D9 reacts with CD55 or DAF (Decay Accelerating Factor). All leucocytes as well as human erythrocytes, fibroblasts, platelets, endothelial cells and neuroectodermal cells are positive for DAF. F4-29D9 also recognizes an antigen on spermatozoid cells. It is a glycosylphosphatidylinositol anchored (GPI-anchored) member of the membrane bound complement regulatory proteins that inhibit autologous complement cascade activation. CD55 also serves as receptor for CD97 and for echovirus and coxsackie B virus. DAF is deficient in both granulocytes and monocytes in patients with paroxysmal nocturnal haemoglobinuria.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
F4-29D9
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kishimoto T. et al., eds. Leukocyte Typing VI, p509-514, Garland Publishing, Inc, (1997)
References 2:
B.E. Loveland - in Leucocyte Typing VI - Part 6 CD55 Workshop Panel Report pp519-520, (1997)
References 3:
Ruix-Delgado, GJ et al, Hematology 14: 33-7 (2009)
F4-31C2 reacts with CD54 or ICAM1 (Intercellular Adhesion Molecule 1). ICAM1 belongs to the immunoglobulin superfamily, C2 subset, is a transmembrane molecule of 90 kDa with 7 potential N-glycosylation sites. It is expressed on resting monocytes and endothelial cells and in response to inflammatory cytokines such as TNF-alpha, IL1 and IFN-gamma, can be highly upregulated on many other cells, e.g. on B- and T-lymphocytes, thymocytes, dendritic cells and also on keratinocytes, chondrocytes, as well as epithelial cells. CD54 mediates cell adhesion by binding to integrin CD11a/CD18 (LFA 1) and to CD1b/CD18 (Mac 1). The interaction of CD54 with LFA 1 enhances antigen specific T-cell activation. CD54 also binds to CD43, fibrinogen, most human rhinoviruses and to Plasmodium falciparum infected erythrocytes. ICAM1 may also be related to progression and metastasis of tumors.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
F4-31C2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Johnson, J.P., et al., in Knapp, W., et al. Leucocyte Typing IV, pp 681-683 (1989)
CD53 is a 33-55 kDa protein expressed on monocytes and macrophages, dendritic cells, osteoblasts and osteoclasts, and on B- and T-cells from every stage of differentiation but is absent from platelets, red blood cells. CD53 appears to be the marker with widest reactivity as well as the marker with the strictest specificity to hematopoietic cells. 161-2 Partially inhibits T-cell proliferation induced by CD3 UCHT-1 antibody. 161-2 was typed in Kobe, Japan at the VIth International Workshop on human leucocyte differentiation antigens.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
161-2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kishimoto T. et al., eds. Leukocyte Typing VI, p509-514, Garland Publishing, Inc, (1997)
101-1D2 reacts with the cellular adhesion molecule CD50 (ICAM-3), a single chain polypeptide with a MW of 12 kDa. The protein is heavily glycosylated and resistant to phosphatidylinositol phospholipase C treatment so probably not PO-anchored. 101-1D2 was provisionally clustered as CDw50 at the IV and confirmed as CD50 at the V international workshop on human leucocyte differentiation antigens.
Cris-1 reacts with a 67 kDa protein, consistent with human CD5. Cris-1 was assigned at the Ist and IIIrd International Leucocyte Typing Workshops. CD5 is a pan T-cell marker that also reacts with a Bcell marker subset and a range of neoplastic B-cells, e.g. chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma, and a subset (~10%) of diffuse large Bcell lymphoma. CD5 aberrant expression is useful in the diagnosis of mature T-cell neoplasms.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
Cris-1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Proceedings of the first international workshop and conference on human leukocyte differentiation antigens. Oxford University Press, Oxford (1983).
References 2:
Leukocyte Typing III, McMichael A. J. et al. (Eds.), Oxford University Press, Oxford (1987)
References 3:
Alberola-Ila J, et al, J Immunol. 148(5): 1287-93 (1992)
References 4:
Arrizabalaga P. et al, Nephron 53: 41-49 (1989)
References 5:
Guarne A. et al, Protein Science 5: 167-169 (1996)
EBS-CD-028 reacts with CD46 or Membrane Cofactor Protein (MCP; 52-74 kDa) All nucleated human cells carry CD46, which has multiple common protein isoforms of 52 kDa to 66 kDa, and 74 kDa on placental cells and some transformed cells. CD46 acts as a cofactor for complement factor I, a serine protease, which protects autologous cell against complement-mediated injury by cleaving C3b and C4b deposited on host tissue. It may further be involved in the fusion of the spermatozoa with the oocyte during fertilization. CD46 acts as a co-stimulatory factor for T-cells, which induces the differentiation of CD4+ into T-regulatory 1 cells. T-regulatory 1 cells suppress immune responses by secreting interleukin-10, and therefore are thought to prevent autoimmunity. A number of viral and bacterial pathogens seem to exploit this property and directly induce an immunosuppressive phenotype in T-cells by binding to CD46.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-028
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Iwata, K., et al. J. Biol. Chem. 270: 15148-15152 (1995)
References 2:
Liszewski, M.K., et al. Adv. Immunol. 61: 201-283 (1996)
References 3:
Liszewski, M.K., et al. J. Immunol. 156: 4415-4421 (1996)
CD45 glycoprotein have various molecular weight on various cell types: B-cells 240 kDa, thymocytes 180 kDa, T-cells multiple bands. Reduced in PAGE gels: 180 and 240 kDa. Isoforms are produced by alternative splicing of domains 4, 5 and 6. Various isoform are expressed differently on different lymphocytes. All hematopoietic cells express CD45 proteins except erythrocytes. Relevant epitopes are termed CD45RA (exon 4), CD45RB (exon 5), CD45RC (exon 6) and CD45R or CD45R0 (exon 4-6 spliced out).
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
Bra55
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Chorváth et al., Neoplasma 34(6), 685-692, (1987)
References 2:
Chorváth et al., Neoplasma 35(5), 495-501, (1988
References 3:
Chorváth et al. Leukocyte Typing IV, pp. 634-637, (1989)
145-23 reacts with CD44, a type 1 transmembrane glycoprotein providing cell-cell and cellmatrix adhesion while linked to the cytoskeleton. It is involved in haematopoiesis, lymphocyte homing and activation, and tumor metastasis. It binds to fibrin, hyaluronate and other matrix proteins. On tumors CD44H is highly expressed, suggesting an important role in progression for this isoform. CD44 also forms the protein backbone of the human erythrocyte Lutheran antigen system. The epitope of 145-23 is resistant to (chemo)trypsin digestion.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
145-23
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Nishina S. et al., Graefes Archiv. Clin. Exp. Opthalm. 235: 92-96 (1997)
References 2:
Horny H.P. et al. Virchows Arch. 429: 91-94 (1996)
84-3C1 reacts with a 95/115 kDa protein on T-cells and thymocytes and a 115/135 kDa molecule on neutrophils and platelets. 70-90% of T-cell lymphomas and from 22-37% of Bcell lymphomas express CD43. No reactivity has been observed with reactive B-cells. So a Blineage population that co-expresses CD43 is highly likely to be a malignant lymphoma, especially a low-grade lymphoma, rather than a reactive B-cell population. When CD43 antibody is used in combination with anti-CD20, effective immunophenotyping of the lymphomas in formalin-fixed tissues can be obtained. Co-staining of a lymphoid infiltrate with anti-CD20 and anti-CD43 argues against a reactive process and favors a diagnosis of lymphoma. In addition, expression is altered in Wiskott Aldrich syndrome. A proportion of AIDS patients have antibodies to CD43. A soluble form called galactoglycoprotein is present in serum. 85-3C1 was typed at the 3rd International Workshop on Human Leucocyte Differentiation antigens.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
84-3C1
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Cobbold, S. et al. In Leucocyte typing III, Oxford University Press, pp 789-803 (1987)
References 2:
Stross, WP. et al. J. Clin. Path. 42: 953-961 (1989)
EDU-2 reacts with human CD4, a 55 kDa single chain transmembrane glycoprotein that contains four extracellular immunoglobulin-like domains and is mainly expressed on T-helper lymphocytes, but also on cortical thymus cells, microglial and dendritic cells. It binds to HLA class II molecules during the interaction of CD4+ T cells with antigen-presenting cells or target cells. CD4 also serves as the primary cellular receptor for human immunodeficiency virus (HIV). EDU-2 was assigned to CD4 at the International Leucocyte Typing Workshops II and IV.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EDU-2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. Leukocyte Typing IV, Oxford Univ. Press, (1989)
References 2:
Reinberz EL et al. eds. Leykcocyte Typing II, Springer-Verlag, Berlin, (1985)
EBS-CD-025 reacts with a 45 kDa glycopeptide which is a type II membrane glycoprotein with a transmembrane sequence near the regulation of lymphocyte adhesion to endothelial cells. Its ligand is CD31. CD38 is found on early cells of B and T lineages and on activated B- and tcells. It is not found on most mature resting peripheral lymphocytes. Also positive are thymus cells and Ig secreting plasma cells. The CD34+ and CD38- population of hematopoietic stems cells defines the most pluripotent cells (e.g. blast colony forming cells). EBS-CD-025 antibody blocks the EBS-CD-026 epitope, and vice versa.
CD33 is a 67 kDa glycoprotein of the sialic acid-binding immunoglobulin-like lectin (SIGLEC) family, displaying the immune-receptor tyrosine-based inhibitory motif (ITIM), able of recruiting protein tyrosine phosphatases SHP-1 and SHP-2 to signal assemblies. ITIMs are also used for ubiquitinmediated removal of the receptor from the cell surface. CD33 is expressed on cells of myelomonocytic lineage, binds sialic acid residues in N- and O-glycans on cell surfaces, and is a therapeutic target for acute myeloid leukemia. CD33 is expressed on myeloid progenitors, monocytes, granulocytes, dendritic cells and mast cells. It is absent on platelets, lymphocytes, erythrocytes and hematopoietic stem cells.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-022
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. Leukocyte Typing IV, Oxford Univ. Press, (1989)
References 2:
Favaloro EJ et al, Dis Markers 5(4): 215-25 (1987)
References 3:
Favaloro EJ et al, Br J Haematol 69(2): 163-71 (1988).
FS12 reacts with CD31 (PECAM-1), a 130-140 kDa member of the immunoglobulin gene superfamily that is expressed on cells of the vasculature, including platelets, endothelial cells, myeloid cells and certain lymphocyte subsets.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
FS12
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Sandilands, G. et al. Clin. and Exp. Immunology, 162(3): 516-27 (2010)
References 2:
Kishimoto T. et al., eds. Leukocyte Typing VI, Garland Publishing, Inc, (1997)
B-B12 reacts with 5 invariable CD3-chains: CD3y or gp26, CD3d or gp20, CD3e or gp20, CD3f or p16 (homodimer), CD3n or p28. Molecular mass: 25-28, 20 and 16 kDa. The main reactivity is with T cells, including thymocytes, mature T cells and T cell lines.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
B-B12
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Meuer SC et al, Nature 303(5920): 808-810 (1983)
References 2:
Reinherz et al, Cell 30: 715 (1982)
References 3:
Borst et al, J. Biol. Chem. 258: 5135 (1983)
References 4:
Van den Elsen et al, Nature 312 (5993): 413-8 (1984)
EBS-CD-017 reacts with CD25 (55 kDa) which associates as alpha chain with CD122 and the common gamma chain (CD132) to form the high-affinity IL-2 receptor complex. With respect to lymphomas, CD25 is present on malignant cells of Hodgkins disease, HTLV-1+ adult T-cell leukemia, cutaneous T-cell lymphoma, and hairy cell leukemia. Increased levels of soluble CD25 are observed in leukemias/lymphomas and inflammatory/ autoimmune diseases. CD25 alone appears to function as a low affinity receptor and associates with CD122 (IL-2R chain, p75) and CD132 (common chain) to form the high affinity IL-2 receptor complex. CD25 antibodies detect three epitope regions, A, B and C. EBS-CD-017 recognizes B epitope, which is located at residue 3-104 of CD25 and does not block IL-2 binding to CD25. CD25 antibodies have been used successfully as a carrier for cytotoxic drugs enabling specific delivery to IL-2RA displaying target cells.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-017
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Yamamura T. et al, Eur J Surg 168(1): 49-54 (2002)
References 2:
Lundin K. et al, Anal Biochem 299(1): 92-7 (2001)
References 3:
Raivio E. et al, APMIS 105(2): 108-14 (1997)
References 4:
Boutin B. et al, Neuropediatrics 20(4): 202-6 (1989)
FR10B4 reacts with high affinity to CD22, which is expressed in the cytoplasma of all B-cells, appearing as early as cell-surface CD19 during B-cell development. Its present on the surface of most mature sIg+ B-cells with especially high expression on hairy cell and prolymphocytic leukemia cells. CD22 is a member of the immunoglobulin super-family and acts as an adhesion molecule: BL-CAM. On frozen sections, CD22 is found highly expressed in follicular mantle and marginal zone B-cells, while germinal centre B-cells react relatively weakly.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
FR10B4
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Campana, D., et al.Leucocyte Typing IV, Oxford Univ. Press, (1989), pp 190-192
FR5A10 reacts with CD21, a 140 kDa cell surface molecule which acts as a receptor for EBV, for human complement factor C3d (CR2) and for IFN-alpha. It is a glycoprotein, made up of multiple (n=15) Short Consensus Repeats (S.C.R.) sequences. FR5A10 has been assigned to (S.C.R.) sequences. FR5A10 has been assigned to S.C.R. numbers 5-8. FR5A10 is highly specific to CR2 and shows no cross-reaction with CR1. CD21 is expressed strongly on mature B-cells, follicular dendritic cells and weakly on immature thymocytes and T-lymphocytes. In B-cell ontogeny, CD21 appears after the preB-stage, is maintained during peripheral B-cell development and is lost upon terminal differentiation into plasma cells. CD21 expression is also gradually lost after stimulation of B-cells in vitro.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
FR5A10
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Schlossman SF et al. Leukocyte Typing?V Oxford?University Press:?342-352 (1995)
References 2:
Aubry JP et al. Leukocyte Typing V, p535-536, Oxford University Press, Oxford, (1995)
93-1B3 binds with CD20 which is a 30/33 kDa non-glycosylated transmembrane phosphoprotein with three extensive hydrophobic regions. CD20 is involved in regulation of Bcell activation. It is expressed on the surface of all B-cells beginning at the pro-B phase (CD45R+, CD117+) and progressively increasing in concentration until maturity. Plasma cells are negative. CD20 is retained on many B-cell malignancies. CD20 positive cells are also sometimes found in cases of Hodgkins disease, myeloma, and thymoma. 93-1B3 has been clustered at the IIIrd and Vth HLDA Workshops.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
93-1B3
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Cobbold, S. et al., in leucocyte typing III, Oxford University Press (1987)
References 2:
Schlossman SF et al. Leukocyte Typing?V Oxford?University Press:?342-352 (1995)
109-3C2 binds with CD20 which is a 30/33 kDa non-glycosylated transmembrane phosphoprotein with three extensive hydrophobic regions. CD20 is involved in regulation of B-cell activation. It is expressed on the surface of all B-cells beginning at the pro-B phase (CD45R+, CD117+) and progressively increasing in concentration until maturity. Plasma cells are negative. CD20 is retained on many B-cell malignancies. CD20 positive cells are also sometimes found in cases of Hodgkins disease, myeloma, and thymoma. 109-3C2 has been clustered at IVth and Vth HLDA Workshops.
Antibody Isotype:
IgG3-K
Monosan Range:
MONOSAN
Clone:
109-3C2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. Leukocyte Typing IV, Oxford Univ. Press, (1989)
References 2:
Schlossman SF et al. Leukocyte Typing?V Oxford?University Press:?342-352 (1995)
CD2 is a transmembrane glycoprotein that is expressed on peripheral blood T lymphocytes, NK cells and thymocytes. Interaction between CD2 and its counter receptor LFA3 (CD58) on opposing cells optimizes immune system recognition, thereby facilitating communication between helper T lymphocytes and antigen presenting cells, as well as between cytolytic effectors and target cells. EBSCD-003 reacts with human T-cells, leukemic T-cells and T-cell lines. EBS-CD-003 also reacts with some, if not all, E-RFC-receptors on K and NK cells.
Antibody Isotype:
IgG2b-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-003
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Thurlow PJ, et al., transplantation 36: 293-298 (1983)
References 2:
Kozarsky KF, et al, Cell Immunol 150: 235-246 (1993)
References 3:
Schlossman SF et al. Leukocyte Typing?V Oxford?University Press:?342-352 (1995)
RIV12 reacts with CD1b, a 44KDa type 1 glycoprotein associated with beta2-microglobulin. It is expressed on dendritic cells, Langerhans cells, thymocytes and T acute lymphoblastic leukemia cells. CD1, type 1 membrane protein, has structural similarity to the MHC class 1 molecule and has been shown to present lipid antigens for recognition by T lymphocytes. CD1b is also expressed in Langerhans interdigitating cells.
CB19 is specific for the antigen CD19. This antigen has a MW of 120 kDa and contains a 280 residue extracellular domain and a 240 residue cytoplasmic domain. It is a critical signal transduction molecule that regulates B-lymphocyte development, activation, and differentiation. It plays a dominant role in establishing signalling thresholds for antigen receptors and other surface receptors on B-lymphocytes. This antigen is lost upon terminal differentiation to plasma cells.
It recognizes a transmembrane glycoprotein of 95 kDa, identified as CD18 or intergrin-2 (Workshop III). It complexes non-covalently with either L, M, or X integrin (CD11a, b, or c) to form the heterodimers. LFA-1, MAC-1, and p150,95, respectively. LFA-1 is the receptor for three members of the Ig supergene family of proteins, ICAM-1 (CD54, ICCAM-2 (CD102), and Mac-1 and p150,95 bind to ICAM-1, fibrinogen, and IC3b. ICAM-3 (CD50). CD18/CD11 heterodimeric molecules are involved with cell/cell and cell/extracellular adhesion in immune and inflammatory responses. This Mab blocks these cellular interactions. 68-5A5 was clustered at the IIIrd International Workshop on human leucocyte differentiation antigens.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
68-5A5
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
McMichael A.J.et al., Leucocyte typing III, Oxford University Press, Oxford, (1987)
CDw17 is an intermediate glycosphingolipid from the metabolism of higher gangliosides that localizes to sphingolipid-sterol rafts. CDw17 is found on monocytes, granulocytes, basophils, platelets, a subset of peripheral B-cells (CD19+) and tonsil dendritic cells. It is rapidly down regulated on activated granulocytes and is upregulated on IL-2 activated T-lymphocytes. CDw17 binds to bacteria and may function in phagocytosis. It may also be involved in angiogenesis. Aberrant levels of glycosphingolipids are a feature of cancer cells and may influence integrin clustering and internalization.
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-014
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Knapp W. Leukocyte Typing IV, Oxford Univ. Press, (1989)
CB16 specifically recognizes CD 16. This molecule also named low affinity Fc-receptor for IgG (FcgammaRIII) exhibits two truncated Ig-like domains and is 50-80 kDa. It is highly expressed on NK-cells, granulocytes and macrophages. CB16 binds to 15% peripheral lymphocytes of healthy donors (NK-cells), granulocytes, macrophages. CD16 represents the functional receptor structure for antibody-dependent cellular cytotoxicity
Antibody Isotype:
IgM-K
Monosan Range:
MONOSAN
Clone:
CB16
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Deaglio S. et al., Blood 99(7): 2490-8 (2002)
References 2:
Zilber MT et al. Proc Natl Acad Sci U S A 97(6): 2840-5 (2000)
References 3:
Wirthmueller U et al. J Exp Med. 175(5): 1381-90 (1992)
EBS-CD-013 specifically recognizes CD16. This molecule also named low affinity Fc-receptor for IgG (FcgammaRIII) exhibits two truncated Ig-like domains and is 50-80 kDa. It is highly expressed on NK-cells, granulocytes and macrophages. EBS-CD-013 binds to 15% peripheral lymphocytes of healthy donors (NK-cells), granulocytes, macrophages. CD16 represents the functional receptor structure for antibody-dependent cellular cytotoxicity.
Antibody Isotype:
IgG2a-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-013
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Deaglio S. et al., Blood 99(7): 2490-8 (2002)
References 2:
Zilber MT et al. Proc Natl Acad Sci U S A 97(6): 2840-5 (2000)
References 3:
Wirthmueller U et al. J Exp Med. 175(5): 1381-90 (1992)
EBS-CD-12 recognizes an extracellular epitope on an integral membrane glycoprotein of 150 kDa, identified as CD13 (also known as aminopeptidase-N). CD13 is present on most cells of myeloid origin including granulocytes, monocytes, mast cells, and GM-progenitor cells. It is also expressed by the majority of AML, CML in myeloid blast crisis, and in a smaller fraction of lymphoid leukemias. CD13 is also present on fibroblasts; endothelial cells, epithelial cells from renal proximal tubules and intestinal brush border, bone marrow stromal cells, osteoclasts, and cells lining bile duct canaliculi. CD13 plays a role in metabolism of biologically active peptides, in phagocytosis, and in bactericidal/tumoricidal activities. It also serves as a receptor for human coronaviruses (hCoV) and human cytomegalovirus (hCMV).
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-012
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Favaloro EJ, et al., Exp Hematol. 13:1695-701 (1993)
References 2:
Favaloro EJ, et al., Clin Chim Acta.220(1):81-90 (1993)
References 3:
Lachance C., et al. J Virol. 72(8):6511-9. (1998)
References 4:
Koch AE, et. al. Am J Pathol. 138(1): 165-73.( 1991)
References 5:
Principe, S. et al, Proteome Res. 6;11(4): 2386-96 (2012)
Integrin ?X (CD11c, leukocyte surface antigen p150/95, CR4, Axb2) is a type 1 transmembrane protein that traditionally combines with ?2 chain to form a leukocyte-specific integrin known as inactivated-C3b (iC3b) receptor 4 (CR4). Integrin ?X/?2 shares similar properties of the Integrin ?M/?2 in mediating adherence of neutrophils and monocytes to stimulated endothelial cells and in phagocytosis of complement coated particles. Abnormal expression of Integrin ?X is characteristic of hairy cell leukemia (HCL) and is dependent upon activation of proto-oncogenes Ras and JunD. Integrin ?x is present on dendritic cells, macrophages and NK-cells. Upon activation, DCs present in skin (Langerhans cells_, lining of nose, lung, stomach, intestine and blood can migrate to lymphoid tissues and interact with T and B-cells to initiate and shape the immune response.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-011
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Cabañas C, et al., Hybridoma 7(2):167-76 (1988)
References 2:
Cabañas C, et al., Immunol Lett. 20(3):193-76 (1988)
References 3:
Zhou JQ, et al. Blood 82:800-6 (1993)
References 4:
Nicolaou, F., et al. Blood 101: 4033-4041 (2003)
References 5:
Edwards, A.D. et al. J. Immunol. 171: 47-60 (2003)
Integrin ?M (also designated complement component receptor 3 ?-chain; CD11b (p170); macrophage antigen ? polypeptide; cell surface glycoprotein Mac-1 ?-subunit; CR3 ?-chain; MAC1A; MO1A and ITGAM) is a cell adhesion molecule that acts as a receptor for cell surface ligands such as intracellular adhesion molecules (ICAMs) or soluble ligands. Integrins are heterodimeric proteins that contain an ?-chain and a ?-chain. Integrin ?M combines with Integrin ?2 (CD18) to form a leukocyte-specific integrin referred to as macrophage receptor-1 (Mac-1) or inactivated-C3b (iC3b) receptor 3 (CR3). Integrin ?M-?2 is important in the adherence of neutrophils and monocytes to stimulated endothelium, and also in the phagocytosis of complement coated particles.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
EBS-CD-010
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Beekhuizen H, et al., J Immunol. 145(2):510-8 (1990)
References 2:
Argenbright LW et al., J Leukoc Biol. 49(3):253-7 (1991)
References 3:
Zhou L, et al. J Biol Chem. 269(25):17075-9 (1994)
References 4:
Miller LJ, et al. J Immunol. 137(9):2891-900 (1986)
FR4D11 reacts with high affinity to CD10 or CALLA, a cell surface enzyme with neutral metalloendopeptidase activity, inactivating a variety of biologically active peptides. CD10 is a 100 kDa glycoprotein, expressed on 70% of pre-B ALL-cells (common ALL), but also on early lymphoid progenitor-cells in bone marrow and fetal liver. Other normal CD10 positive tissues include renal epithelium, fibroblasts and germinal centre B-cells. Density of CD10 antigen has been shown to be related to cell differentiation and may have prognostic value for B-cell lineage acute leukemia. CD10 is also present on breast myoepithelial cells, bile canaliculi, fibroblasts, with especially high expression on the brush border of kidney and gut epithelial cells.
Antibody Isotype:
IgG1-K
Monosan Range:
MONOSAN
Clone:
FR4D11
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Brown, B. et al., J. Natl. Canc. Inst,.55: 1281-1289 (1975)
References 2:
Tran-Paterson, R. et al., Blood, 76: 775-782 (1990)
References 3:
Doerken, B. et al., in Knapp, W. et. al. (eds)., Leucocyte Typing IV, Oxford Univ. Press, pp 33-34
Freshly ejaculated human sperms were washed in PBS and extracted in 3% acetic acid, 10% glycerol, 30 mM benzaminidine. The acid extract was dialyzed against 0.2% acetic acid and subsequently used for immunization.
Clusterin (APO J, SGP-2, TRPM-2, SP-40, pADHC-9, CLJ, T64, GP III, XIP8) is a 75-80 kD disulfide-linked heterodimeric protein containing about 30% of N-linked carbohydrate rich in sialic acid but truncated forms targeted to the nucleus have also been identified. It is a conserved secreted glycoprotein expressed by a wide range of tissues and being implicated in many physiological processes, including e.g. lipid transportation, complement inhibition, tissue remodeling, membrane recycling, or clearence of cellular debris. It is nearly ubiqitously expressed in most mammalian tissues and can be found in plasma, milk, urine, cerebrospinal fluid and semen. Clusterin is able to bind and form complexes with numerous partners (immunoglobulins, lipids, heparin, bacteria, complement components, paraoxonase, beta amyloid, leptin etc.) and is expressed in many pathological and clinically relevant situations including cancer, organ regeneration, infection, Alzheimer disease, retinitis pigmentosa, myocardial infarction, renal tubular damage, autoimmunity and others. A genuine function of clusterin is still enigmatic.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
Hs-3
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Lyn is a Src-family protein tyrosine kinase that is predominantly expressed in hematopoietic cells. It is associated with a number of cell surface receptors including the B cell antigen receptor (BCR) and Fc receptors. Upon their triggering, Lyn phosphorylates subunits of these receptors in a cholesterol-dependent manner, utilizing the plasma membrane lipid raft system. The phosphorylated intracellular domains of the receptors are accessible for cytoplasmic Syk tyrosine kinase, which is activated by Lyn-mediated phosphorylation and which transduces the signal to downstream adaptors. Lyn is abnormally distributed in acute myeloid leukemia cells and seems to be a novel pharmacologic target of this disease.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
LYN-01
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Fyn is a ubiquitously expressed Src-family protein tyrosine kinase with important roles e.g. in immune and nervous system. It regulates N-methyl-D-aspartate (NMDA) receptor functions, thus affecting various brain functions, and even many of its other substrates are important for neural migration, synaptic plasticity, oligodendrocyte differentiation, and axon growth and guidance. In immune system Fyn namely regulates the commitment of T cells to activation, is important in T cell anergy induction, promotes mast cell chemotaxis and reorganization of cytoskeleton and participates in mast cell activation. Fyn is also involved in embryonic stem cell growth and differentiation, associates with tubulin and may play roles in mitotic spindle formation.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
FYN-1
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
AE-2 recognizes double stranded DNA expressed in the nucleus of eukaryotes. Dilution advice: Flow cytometry (1-2 µg/million cells in 0,1 ml, fix cells in 4% PFA for 10 min, at 4°C, permeabilize with 0,2% saponin or digitonin for 15 min, at 4°C). ? Immunofluorescence (1-2 µg/ml). ? Immunohistology (1-2 µg/ml for 30 min at RT; staining of formalin-fixed tissues requires incubating tissue sections in 4N HCl, for 30 minutes).
Antibody Isotype:
IgG3
Monosan Range:
MONOSAN
Clone:
AE-2
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Epstein, AL et al. In Progress on nonhistone protein research. 1: 117-137 (1987)
SOCS3 (suppressor of cytokine signaling 3), also known as CIS3 (cytokine-inducible SH2 protein 3) is a negative regulator of particular cytokine signaling pathways. SOCS3 is induced by a variety of cytokines and other stimuli, such as erythropoietin, leptin and lipopolysaccharides and inhibits tyrosinkinase activity of JAK kinases, or e.g. JNK phosphorylation. SOCS3 modulates cytokine-mediated and neoplastic-proliferative responses and is involved also in maintaining leukocytes in quiescent state until antigen stimulation.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
SO1
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The monoclonal antibody K5A6 recognizes human liver fatty acid binding protein (L-FABP) of both natural and recombinant origin. The L-FABP protein is derived from the human FABP1 gene. FABPs are small intracellular proteins (~13-14 kDa) with a high degree of tissue specificity that bind long chain fatty acids. They are abundantly present in various cell types and play an important role in the intracellular utilization of fatty acids, transport and metabolism. There are at least nine distinct types of FABP, each showing a specific pattern of tissue expression. Due to its small size, FABP leaks rapidly out of ischemically damaged necrotic cells leading to a rise in serum levels. Ischemically damaged tissues are characterized histologically by absence (or low presence) of FABP facilitating recognition of such areas. L-FABP is localized in the liver, kidney and intestinal epithelium. The monoclonal antibody K5A6 is useful to detect ischemic areas of human liver.
The monoclonal antibody L2B10 recognizes human liver fatty acid binding protein (L-FABP) of both natural and recombinant origin. The L-FABP protein is derived from the human FABP1 gene. FABPs are small intracellular proteins (~13-14 kDa) with a high degree of tissue specificity that bind long chain fatty acids. They are abundantly present in various cell types and play an important role in the intracellular utilization of fatty acids, transport and metabolism. There are at least nine distinct types of FABP, each showing a specific pattern of tissue expression. Due to its small size, FABP leaks rapidly out of ischemically damaged necrotic cells leading to a rise in serum levels. Ischemically damaged tissues are characterized histologically by absence (or low presence) of FABP facilitating recognition of such areas. L-FABP is localized in the liver, kidney and intestinal epithelium. The monoclonal antibody L2B10 is useful to detect ischemic areas of human liver. Furthermore, the antibody can be used for the purification of human L-FABP.
Antibody Isotype:
IgG2b
Monosan Range:
MONOSAN
Clone:
L2B10
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Bax; D et al. Scand J Gastroenterology 2007; 42: 902
The monoclonal antibody 1G11B1 recognizes vascular cell adhesion molecule-1 (VCAM-1). VCAM-1 is a member of the immunoglobulin superfamily of adhesion molecules, which includes ICAMs, PECAM-s and MADCAM, and is involved in leukocyte-endothelial cell interactions. The immunoglobulin superfamily is a type I transmembrane protein characterized by extracellular immunoglobin domains, a transmembrane region and a cytoplasmic tail. They are essential for the development of the embryo and for immune and inflammatory responses. These transmembrane glycoproteins mediate cell interaction with, and adhesion to, other cells and the extracellular matrix. VCAM-1 contains six immunoglobulin domains of the H-type and interacts with VLA-4 expressed on leukocytes. Multiple adhesion molecules play a role in leukocyte recruitment. The process of migration of a leukocyte through the vascular endothelium consists of the following steps: leukocyte-endothelium interaction (first tethering and rolling and than adhesion) and transendothelial migration. VCAM-1 is almost not expressed under physiological conditions. However, under appropriate pro-inflammatory conditions where the endothelium is exposed to inflammatory cytokines such as tumour necrosis factor-? or IL-1b and becomes activated, VCAM-1 gene expression is rapid elevated by the vascular endothelium. There is also a soluble form of VCAM-1 which is angiogenic and chemotactic for endothelial cells. sVCAM-1 is up-regulated in several disease states (eg, myocardial infarction, type 2 diabetes mellitus, primary antiphospholipid syndrome, and rheumatoid arthritis).
ENA2 reacts with E-selectin CD62-E, previous designated the Endothelial Leucocyte Adhesion Molecule-1 (ELAM-1). The antibody reacts with human endothelial cells activated with TNF-alpha, IL-1 or endotoxin. The antibody was found to react also with cells transfected with the E-selectin gene. The antibody inhibits the adhesion of granulocytes both neutrophilic and eosinophilic.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
ENA2
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Leeuwenberg; JFM et al. Eur J Immunol 1989; 19: 715
References 2:
Leeuwenberg, JFM et al Clin and Exp Immunol 1990, 81: 496
References 3:
Leeuwenberg; JFM et al. Immunology 1990; 71: 301
References 4:
Leeuwenberg JFM et al. J Immunology 1990; 145: 2110
ENA1 reacts with E-selectin CD62-E, previous designated the Endothelial Leucocyte Adhesion Molecule-1 (ELAM-1). The antibody reacts with human endothelial cells activated with TNF-alpha, IL-1 or endotoxin. The antibody was found to react also with cells transfected with the E-selectin gene. The antibody inhibits the adhesion of granulocytes both neutrophilic and eosinophilic.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
ENA1
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Leeuwenberg; JFM et al. Eur J Immunol 1989; 19: 715
References 2:
Leeuwenberg, JFM et al Clin and Exp Immunol 1990, 81: 496
References 3:
Leeuwenberg; JFM et al. Immunology 1990; 71: 301
References 4:
Leeuwenberg JFM et al. J Immunology 1990; 145: 2110
The antibody binds to the 10 kD eosinophil Major Basic Protein (MBP) of both resting and activated eosinophils in cytospins and frozen sections of bronchial and skin biopsies of allergic sites and normal sites and thus can be used as a "pan-eosinophil" marker. The antibody BMK-13 stains in frozen sections of bronchial biopsies from atopic asthmatics, rhinitics and normal non-atopic subjects, substantially higher counts of positive cells when compared to EG1, EG2 and chromotrope 2R. BMK-13 cross-reacts weakly with human basophils, which also contain low level of this protein. It does not cross-react with any other human protein or cell.
This antibody stains all human blood vessels including brain microvessels. Both large and small vessels are equally reactive. The EN4 antigen is preserved in endothelial cells in culture unlike the PAL-E antigen which is lost. It stains strongly murine fibroblasts transfected with the human CD31 gene. On surface-iodinated Jurkat T cells it recognizes the 130 kD CD31 antigen. EN4 also binds to guinea-pig and cat endothelium, but not to rabbit, bovine, sheep, dog, rat or mice endothelial cells. No other structures in the skin, heart, kidney, tonsils or spleen are stained with this antibody.
The antibody only stains human and various animal blood vessels with exception of arteries. It does not stain rat, mouse and chicken endothelium. The antibody is useful for immunohistochemistry on acetone fixed frozen sections. Not useful on cellsuspensions and Western blotting.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
PAL-E
Concentration:
15 ug/ml
Storage buffer:
Culture medium with 0.01M Sodium Azide
Storage:
2-8°C
References 1:
Jones, R.R., et al., J. Clin. Path. 39, 742-749 (1986).
References 2:
Holden, C.A., et al., J. Immun. Meth. 91, 45-52 (1986).
References 3:
Schlingemann, R.O., et al., Laboratory Investigation 52, 71 (1987).
CD22 protein may be involved in the localization of B-cells in lymphoid tissues. CD22 is expressed in the cytoplasm and cell membrane of B-cells. CD22 is especially useful in diagnostics of hairy cell leukemia and classification of the B-cell lymphomas.
Vimentin is the major subunit protein of the intermediate filaments of mesenchymal cells. It is believed to be involved with the intracellular transport of proteins between the nucleus and plasma membrane. Vimentin has been implicated to be involved in the rate of steroid synthesis via its role as a storage network for steroidogenic cholesterol containing lipid droplets. Vimentin phosphorylation by a protein kinase causes the breakdown of intermediate filaments and activation of an ATP and myosin light chain dependent contractile event. This results in cytoskeletal changes that facilitate the interaction of the lipid droplets within mitochondria, and subsequent transport of cholesterol to the organelles leading to an increase in steroid synthesis. Immunohistochemical staining for Vimentin is characteristic of sarcomas (of neural, muscle and fibroblast origin) compared to carcinomas which are generally negative. Melanomas, lymphomas and vascular tumors may all stain for Vimentin. Vimentin antibodies are thus of value in the differential diagnosis of undifferentiated neoplasms and malignant tumors. They are generally used with a panel of other antibodies including those recognising cytokeratins, lymphoid markers, S100, desmin and neurofilaments.
CD14 antigen is a GPI-linked glycoprotein with a molecular weight of 55kD. The CD14 antigen is expressed on cells of the myelomonocytic lineage including monocytes, macrophages and Langerhans cells. Low expression is observed on neutrophils and on human B cells. CD14 antigen is a receptor for bacterial lipopolysaccharide (LPS, endotoxin) and the lipopolysaccharide binding protein (LBP). LBP and CD14 antigen serves two physiological roles. These proteins act as opsonin and opsonic receptor, respectively, to promote the phagocytic uptake of bacteria or LPScoated particles by macrophages.
The androgen receptor (AR), also known as NR3C4 (nuclear receptor subfamily 3, group C, member 4), is a type of nuclear receptor which is activated by binding of either of the androgenic hormones testosterone or dihydrotestosterone in the cytoplasm and then translocating into the nucleus. The androgen receptor is most closely related to the progesterone receptor, and progestins in higher dosages can block the androgen receptor. The main function of the androgen receptor is as a DNA binding transcription factor which regulates gene expression; however, the androgen receptor has other functions as well. Androgen regulated genes are critical for the development and maintenance of the male sexual phenotype.
AMACR (alpha-methylacyl-CoA racemase) has been recently described as prostate cancer-specific gene that encodes a protein involved in the beta-oxidation of branched chain fatty acids. Expression of AMACR protein is found in prostatic adenocarcinoma but not in benign prostatic tissue. It stains premalignant lesions of prostate: high-grade prostatic intraepithelial neoplasia (PIN) and atypical adenomatous hyperplasia. AMACR can be used as a positive marker for PIN. Defects in AMACR are the cause of congenital bile acid synthesis defect type 4 (CBAS4); also known as cholestasis, intrahepatic, with defective conversion of trihydroxycoprostanic acid to cholic acid or trihydroxycoprostanic acid in bile. Clinical features include neonatal jaundice, intrahepatic cholestasis, bile duct deficiency and absence of cholic acid from bile.
Cytokeratins are classified into one of two classes, type I (acidic polypeptides) and type II (basic polypeptides). Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. Cytokeratins have been found to be useful markers of tissue differentiation which is directly applicable to the characterization of malignant carcinomas.
Ki67, also known as MKI67, is aprototypic cell cycle related nuclear protein, expressed by proliferating cells in all phases of the active cell cycle (G1, S, G2 and M phase). It is absent in resting (G0) cells. Ki67 antibodies are useful in establishing the cell growing fraction in neoplasms (immunohistochemically quantified by determining the number of Ki67 positive cells among the total number of resting cells = Ki67 index). In neoplastic tissues the prognostic value is comparable to the tritiated thymidine labelling index. The correlation between low Ki67 index and histologically low grade tumours is strong. Ki67 is routinely used as a neuronal marker of cell cycling and proliferation
ERBB2: v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian). This gene encodes a member of the epidermal growth factor (EGF) receptor family of receptor tyrosine kinases. This protein has no ligand binding domain of its own and therefore cannot bind growth factors. However, it does bind tightly to other ligand-bound EGF receptor family members to form a heterodimer, stabilizing ligand binding and enhancing kinase-mediated activation of downstream signalling pathways, such as those involving mitogen-activated protein kinase and phosphatidylinositol-3 kinase. Allelic variations at amino acid positions 654 and 655 of isoform a (positions 624 and 625 of isoform b) have been reported, with the most common allele, Ile654/Ile655, shown here. Amplification and/or overexpression of this gene has been reported in numerous cancers, including breast and ovarian tumors. Alternative splicing results in several additional transcript variants, some encoding different isoforms and others that have not been fully characterized
Glucagon, is a pancreatic hormone that counteracts the glucose-lowering action of insulin by stimulating glycogenolysis and gluconeogenesis. Glucagon is a ligand for a specific G-protein linked receptor whose signalling pathway controls cell proliferation. Two of the other peptides are secreted from gut endocrine cells and promote nutrient absorption through distinct mechanisms. Finally, the fourth peptide is similar to glicentin, an active enteroglucagon. Glucagon is secreted in the alpha cells of the islets of Langerhans. GLP-1, GLP-2, oxyntomodulin and glicentin are secreted from enteroendocrine cells throughout the gastrointestinal tract. GLP1 and GLP2 are also secreted in selected neurons in the brain.
E-Cadherin is a 120 kDa transmembrane glycoprotein that is localized in the adherens junctions of epithelial cells. There, it interacts with the cytoskeleton through the associated cytoplasmic catenin proteins. In addition to being a calcium-dependent adhesion molecule, E-Cadherin is also a critical regulator of epithelial junction formation. Its association with catenins is necessary for cell-cell adhesion. These E-cadherin/catenin complexes associate with corical actin bundles at both the zonula adherens and the lateral adhesion plaques. Tyrosine phosphorylation can disrupt these complexes, leading to changes in cell adhesion properties. E-Cadherin expression is often down-regulated in highly invasive, poorly differentiated carcinomas. Increased expression of E-Cadherin in these cells reduces invasiveness. Thus, loss of expression or function of E-Cadherin appears to be an important step in tumorigenic progression.Tissue specificity: Non-neural epithelial tissues.
Desmin (DES), with 470-amino acid protein (about 52kDa), belongs to the intermediate filament family and Desmin is class III intermediate filaments found in muscle cells. Homopolymers of Desmin form a stable intracytoplasmic filamentous network connecting myofibrils to each other and to the plasma membrane.Mutations in Desmin are associated with desmin-related myopathy, a familial cardiac and skeletal myopathy (CSM), and with distal myopathies.Desmin is also expressed in smooth muscle cells of both airways and alveolar ducts and Desmin is a load-bearing protein that stiffens the airways and consequently the lung and modulates airway contractile response.
Cytokeratin 19, also known as KRT19, CK19, CK19, K1CS, MGC15366. Entrez Protein NP_002267. It is a member of the keratin family. The keratins are intermediate filament proteins responsible for the structural integrity of epithelial cells and are subdivided into cytokeratins and hair keratins. The type I cytokeratins consist of acidic proteins which are arranged in pairs of heterotypic keratin chains. Unlike its related family members, this smallest known acidic cytokeratin is not paired with a basic cytokeratin in epithelial cells. It is specifically expressed in the periderm, the transiently superficial layer that envelopes the developing epidermis.
Cytokeratin 18, also known as CK18, CYK18, KRT18. Entrez Protein NP_000215. It encodes the type I intermediate filament chain keratin 18. Keratin 18, together with its filament partner keratin 8, are perhaps the most commonly found members of the intermediate filament gene family. They are expressed in single layer epithelial tissues of the body. Mutations in this gene have been linked to cryptogenic cirrhosis. Two transcript variants encoding the same protein have been found for this gene.
CK17, also known as KRT17, it is the type I intermediate filament chain keratin 17. It is found in nail beds, hair follicles, sebaceous glands, and other epidermal appendages. Mutations in this gene lead to Jackson-Lawler type pachyonychia congenita and steatocystoma multiplex. May play a role in the formation and maintenance of various skin appendages, specifically in determining shape and orientation of hair. May be a marker of basal cell differentiation in complex epithelia and therefore indicative of a certain type of epithelial "stem cells". May act as an autoantigen in the immunopathogenesis of psoriasis, with certain peptide regions being a major target for autoreactive T-cells and hence causing their proliferation. Required for the correct growth of hair follicles, in particular for the persistence of the anagen (growth) state. Modulates the function of TNF-alpha in the specific context of hair cycling. Regulates protein synthesis and epithelial cell growth through binding to the adapter protein SFN and by stimulating Akt/mTOR pathway. Involved in tissue repair.
Epithelial Cell Adhesion Molecule (EpCAM) is a 40 kDa cell surface antigen and this protein is expressed in almost all epithelial cell membranes but not on mesodermal or neural cell membranes. EpCAM is a Type 1 transmembrane glycoprotein and it is expressed on the basolateral membrane of cells by the majority of epithelial tissues, with the exception of adult squamous epithelium and some specific epithelial cell types including hepatocytes and gastric epithelial cells. EpCAM expression has been reported to be a possible marker of early malignancy, with expression being increased in tumor cells, and de novo expression being seen in dysplastic squamous epithelium. This cell surface, glycosylated 40kD protein is highly expressed in the bone marrow, colon, lung, and most normal epithelial cells and is expressed on carcinomas of gastrointestinal origin.
CD10 is a 100kDa glycoprotein, also designated Common Acute Lymphocytic Leukemia Antigen (CALLA). It is a cell surface enzyme with neutral metalloendopeptidase activity which inactivates a variety of biologically active peptides. CD10 is expressed on the cells of lymphoblastic, Burkitts, and follicular germinal center lymphomas, and on cells from patients with chronic myelocytic leukemia (CML). It is also expressed on the surface of normal early lymphoid progenitor cells, immature B cells within adult bone marrow and germinal center B cells within lymphoid tissue. CD10 is also present on breast myoepithelial cells, bile canaliculi, fibroblasts, with especially high expression on the brush border of kidney and gut epithelial cells.
Mouse anti Human CD90 antibody, clone F15-42-1 recognizes the human CD90 cell surface antigen, a ~25 kDa glycoprotein homologous to rat Thy1. The antigen is expressed by a subset of CD34+ve cells in the bone marrow and by prothymocytes within the thymus. CD90 is also expressed extensively within the brain.Mouse anti Human CD90 antibody, clone F15-42-1 is routinely tested in flow cytometry on the MOLT4 cell line.
Mouse anti Human CD90 antibody, clone F15-42-1 recognizes the human CD90 cell surface antigen, a ~25 kDa glycoprotein homologous to rat Thy1. The antigen is expressed by a subset of CD34+ve cells in the bone marrow and by prothymocytes within the thymus. CD90 is also expressed extensively within the brain.Mouse anti Human CD90 antibody, clone F15-42-1 is routinely tested in flow cytometry on the MOLT4 cell line.
Mouse anti Human CD90 antibody, clone F15-42-1 recognizes the human CD90 cell surface antigen, a ~25 kDa glycoprotein homologous to rat Thy1. The antigen is expressed by a subset of CD34+ve cells in the bone marrow and by prothymocytes within the thymus. CD90 is also expressed extensively within the brain.Mouse anti Human CD90 antibody, clone F15-42-1 is routinely tested in flow cytometry on the MOLT4 cell line.
Mouse anti Human CD200R antibody, clone OX108 recognizes human CD200R, a cell surface glycoprotein (also known as OX2R). In humans CD200R is expressed primarily by peripheral blood monocytes and neutrophils but also by other leucocytes including T-lymphocytes and mast cells, CD200-CD200R interaction may be involved in the control of myeloid cellular function (Wright et al. 2003). Levels of expression on resting peripheral blood cells are relatively low.
Insulin is a protein consisting of an ?-chain of 21 amino acids and a ?-chain of 30 amino acids and produced in the ?-cells of the pancreas. 2D11-H5 is a specific insulin antibody as tested by ELISA and on human pancreatic tissue.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
2D11-H5
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
De la Tour, D, et al, Mol. Endoc. 15: 476-483 (2001)
References 2:
Rajagopal, J, et al, Science 299: 363 (2003)
References 3:
Morisset, J, et al, J. Histochem. Cytochem. 51: 1501-1513 (2003)
The alpha interferons are involved in virus resistance in target cells for these viruses. They are known to block cell proliferation and to regulate MHC class I antigen expression. The IFN? family has over 20 genes and pseudogenes in two families (I and II), one with a mature length of 166aa and one of 172aa. Cells producing IFN? are lymphocytes, monocytes, macrophages and cell lines such as Namalwa and KGI. Bioassays for IFN? include cytopathic effect blocking, by viruses such as VSV, SFV and BMCV, on their target cells. A number of receptors for IFN? are now known and seem to be expressed on most cell types. N39 is specific for human IFN?2 and does not cross react with human IFN?1. N39 is directed against immunodominant epitope site I (aa112-148).
Antibody Isotype:
IgG1,kappa
Monosan Range:
MONOSAN
Clone:
N39
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kontsek, P. et al., Mol Immunol. 29: 863-870 (1992)
References 2:
Kontsek, P. et al., Immunol. Lett. 35: 281-284 (1993)
Insulin is a protein consisting of an ?-chain of 21 amino acids and a ?-chain of 30 amino acids and produced in the ?-cells of the pancreas. E2-E3 is a specific insulin antibody as tested by ELISA and on human pancreatic tissue.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
E2-E3
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
De la Tour, D, et al, Mol. Endoc. 15: 476-483 (2001)
References 2:
Rajagopal, J, et al, Science 299: 363 (2003)
References 3:
Morisset, J, et al, J. Histochem. Cytochem. 51: 1501-1513 (2003)
The alpha interferons are involved in virus resistance in target cells for these viruses. They are known to block cell proliferation and to regulate MHC class I antigen expression. The IFN? family has over 20 genes and pseudogenes in two families (I and II), one with a mature length of 166aa and one of 172aa. Cells producing IFN? are lymphocytes, monocytes, macrophages and cell lines such as Namalwa and KGI. Bioassays for IFN? include cytopathic effect blocking, by viruses such as VSV, SFV and BMCV, on their target cells. A number of receptors for IFN? are now known and seem to be expressed on most cell types. 2-52 Is specific for human IFN?1 and does not cross react with human IFN?2.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
Feb-52
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kontsek, P. et al., Mol Immunol. 29: 863-870 (1992)
The alpha interferons are involved in virus resistance in target cells for these viruses. They are known to block cell proliferation and to regulate MHC class I antigen expression. The IFN? family has over 20 genes and pseudogenes in two families (I and II), one with a mature length of 166aa and one of 172aa. Cells producing IFN? are lymphocytes, monocytes, macrophages and cell lines such as Namalwa and KGI. Bioassays for IFN? include cytopathic effect blocking, by viruses such as VSV, SFV and BMCV, on their target cells. A number of receptors for IFN? are now known and seem to be expressed on most cell types. 2-48 Is specific for human interferon ? 1 and does not cross react with human interferon ? 2.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
Feb-48
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kontsek, P. et al., Mol Immunol. 29: 863-870 (1992)
The alpha interferons are involved in virus resistance in target cells for these viruses. They are known to block cell proliferation and to regulate MHC class I antigen expression. The IFN? family has over 20 genes and pseudogenes in two families (I and II), one with a mature length of 166aa and one of 172aa. Cells producing IFN? are lymphocytes, monocytes, macrophages and cell lines such as Namalwa and KGI. Bioassays for IFN? include cytopathic effect blocking, by viruses such as VSV, SFV and BMCV, on their target cells. A number of receptors for IFN? are now known and seem to be expressed on most cell types. N27 is specific for human IFN?2 and does not cross react with human IFN?1. N27 reacts with linear peptide 43aa-53aa, placing the epitope outside the immunodominant regions I and II.
Antibody Isotype:
IgG1,kappa
Monosan Range:
MONOSAN
Clone:
N27
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Kontsek, P. et al., Mol Immunol. 29: 863-870 (1992)
References 2:
Kontsek, P. et al., Immunol. Lett. 35: 281-284 (1993)
The antibody reacts specificly with Human IL-1RII. The IL-1 system includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1RI and IL-1RII) and the IL-1 receptor accessory protein. The IL-1RII is part of the antagonistic IL-1 mechanism. It is also known as decoy receptor and is a non signaling molecule which functions by capturing IL-1 and preventing it from interacting with the signalling IL-1RI. The decoy IL-1RII can after binding to IL-1 also recruit the IL-1 receptor accessory protein and thus inhibit by coreceptor competition. Further a soluble form of IL-1RII exists which is shed, a process in which matrix metalloproteases have been found to play a role, by various cells including monocytes, polymorphonuclear cells, B cells and fibroblasts.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
6G5
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Mantovani; A et al. Ann N Y Acad Sci 1998; 840: 338
Mouse gamma light chain of Immunoglobulin (determined by immunodot) and exhibits no reactivity to mouse lambda light chain. No detectable reactivity with human IgG as tested by ELISA. Available in unconjugated MON5083, FITC conjugated MON5083F, HRP conjugated MON5063H and Biotin conjugated MON5063B.
Mouse gamma light chain of Immunoglobulin (determined by immunodot) and exhibits no reactivity to mouse lambda light chain. No detectable reactivity with human IgG as tested by ELISA. Available in unconjugated MON5083, FITC conjugated MON5083F, HRP conjugated MON5063H and Biotin conjugated MON5063B.
Monoclonal antibody binds both natural and recombinant human gamma Interferon. Cross reactivity with other cytokines has not been found. The antibody does not react with rodent interferons or with alpha or beta interferons.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
F14
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Meide van der; PH et al. J Immunol Methods 1985; 79: 293
The antibody reacts specificly with Human IL-1ra3. IL-1ra3 belongs to the IL-1 system which includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1 R I and IL-1 R II) and the IL-1 receptor accessory protein. Three molecular isoforms of the IL-1 receptor antagonist (IL-1ra) have been identified and cloned. Secreted IL-1ra (sIL-1ra or IL-1ra1) contains a classical leader peptide giving a released mature protein. Two intracellular isoforms, icIL-1ra type I (IL-1ra2) and icIL-1 ra type II (IL-1ra3), have no leader sequence, thus predicting that these proteins remain intracellular. IL-1ra3 may represent a reservoir of IL-1 inhibitors, released upon cell death, whose function is to limit the pro-inflammatory action of cell debris. Studies have shown that IL-1ra3 is expressed by various cell types upon exposure to inflammatory signals but most prominently by mononuclear phagocytes and keratinocytes.
The antibody reacts specificly with Human IL-1RII. The IL-1 system includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1RI and IL-1RII) and the IL-1 receptor accessory protein. The IL-1RII is part of the antagonistic IL-1 mechanism. It is also known as decoy receptor and is a non signaling molecule which functions by capturing IL-1 and preventing it from interacting with the signalling IL-1RI. The decoy IL-1RII can after binding to IL-1 also recruit the IL-1 receptor accessory protein and thus inhibit by coreceptor competition. Further a soluble form of IL-1RII exists which is shed, a process in which matrix metalloproteases have been found to play a role, by various cells including monocytes, polymorphonuclear cells, B cells and fibroblasts.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
8.5
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Mantovani; A et al. Ann N Y Acad Sci 1998; 840: 338
The monoclonal antibody binds to soluble mouse interleukin-1 receptor type I. The IL-1 system includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1RI and IL-1RII) and the IL-1 receptor accessory protein. Interleukin-1 signal is transduced through the type I receptor.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
D14e3
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Kojouharoff; G et al. Clin Exp Immunol 1997; 107: 353
The monoclonal antibody binds to soluble mouse interleukin-1 receptor type I. The IL-1 system includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1RI and IL-1RII) and the IL-1 receptor accessory protein. Interleukin-1 signal is transduced through the type I receptor.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
D1f3
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Kojouharoff; G et al. Clin Exp Immunol 1997; 107: 353
The monoclonal antibody binds to soluble mouse interleukin-1 receptor type I. The IL-1 system includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1RI and IL-1RII) and the IL-1 receptor accessory protein. Interleukin-1 signal is transduced through the type I receptor.
Antibody Isotype:
IgG2a
Monosan Range:
MONOSAN
Clone:
Reg20
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Kojouharoff; G et al. Clin Exp Immunol 1997; 107: 353
The monoclonal antibody binds to soluble mouse interleukin-1 receptor type I. The IL-1 system includes two agonists (IL-1alpha and IL-1beta), converting enzymes, antagonists, two receptors (IL-1RI and IL-1RII) and the IL-1 receptor accessory protein. Interleukin-1 signal is transduced through the type I receptor.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
Reg21
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Kojouharoff; G et al. Clin Exp Immunol 1997; 107: 353
The antibody neutralizes mouse interleukin 6 (IL-6). It does not cross-react with LPS (the component of the bacterial cell wall which is considered responsible for the toxicity of gram-negative bacteria) and TNFalpha nor does it bind to either human or rat IL-6. The antibody inhibits IL-6 induced proliferation of the B9 assay. IL-6 is a pluripotent 20-22 kDa cytokine which plays a role in the pathophysiology of severe infection and regulates the immune response, acute phase reaction and haematopoiesis. IL-6 plays a critical role in B-cell differentiation to plasma cells and is a potent growth factor for plasmacytoma and myeloma. Continuous IL-6 gene expression makes an essential contribution to a multistep oncogenesis of plasma cell neoplasia. IL-6 is a very useful culture supplement for the generation of a high number of antibody-producing hybridomas.
Monoclonal antibody F3 binds and neutralizes both natural and recombinant mouse gamma Interferon. Its binding and neutralizing activity has been demonstrated in vitro and in vivo. F3 antibodies have been demonstrated to be able to inhibit inflammatory responses to bacterial lipopolysaccharides. These antibodies were furthermore shown to inhibit Shwartzman reactions and to protect NZB mice against spontaneous development of autoimmune disease. The antibody does not react with rat or human gamma interferon.
Monoclonal antibody F1 binds both natural and recombinant mouse gamma Interferon. Its binding activity has been demonstrated in vitro and in vivo. F1 antibodies have been demonstrated to be able to inhibit inflammatory responses to bacterial lipopolysaccharides. These antibodies were furthermore shown to inhibit Shwartzman reactions and to protect NZB mice against spontaneous development of autoimmune disease. The neutralizing activity of the antibody has been demonstrated as being poor in anti-viral assays. If a neutralizing antibody is specifically desired, we recommend the use of another antibody available from Hycult Biotech, namely F3 (prod. code HM1005) which recognizes a different epitope on the murine gamma Interferon molecule and in contrast to F1, exhibits strong neutralizing activity. The antibody does not react with rat or human gamma interferon
Mouse gamma 2a heavy chain of immunoglobulin (determined by immunodot). No crossreactivity with other animals. Useful in immunoassays, such as ELISA's (good binding on plastic), immunohistochemistry on frozen sections and for affinity chromatog¬raphy of murine IgG1. Available unconjugated MON5068, HRP conjugated 5068H
Mouse gamma 2a heavy chain of immunoglobulin (determined by immunodot). No crossreactivity with other animals. Useful in immunoassays, such as ELISA's (good binding on plastic), immunohistochemistry on frozen sections and for affinity chromatog¬raphy of murine IgG1. Available unconjugated MON5068, HRP conjugated 5068H
The monoclonal antibody F18 recognizes and neutralizes both natural and recombinant mouse alpha Interferon (IFN-?). IFN-? is a cytokine that belongs to the type I interferons (IFN-I). IFN-? is secreted by many cell types including lymphocytes (NK cells, B-cells and T-cells), macrophages, fibroblasts, endothelial cells, osteoblasts, microglia and others. Interferons stimulate both macrophages and NK cells to elicit an anti-viral response, and are also active against tumors. Although all cells can produce IFN-I, plasmacytoid dendritic cells (pDCs) produce 1,000-fold higher levels than other cell types, and are responsible for systemic IFN-I responses to many viruses. They are coined as the natural IFN-producing cells. However, under deprived pDC condition, other dendritic cells are capable of producing high levels of IFN-I.<br /> Interferons were initially characterized for their ability to 'interfere' with viral replication, slow cell proliferation, and profoundly alter immunity. IFN-? has several regulatory roles and diverse biological activities, including control of cellular and humoral immune responses, inflammation, and tumor regression. In addition, IFN-? participates in the regulation of various cellular and humoral processes such as the endocrine system modulates behavior, brain activity, temperature, glucose sensitive neurons, feeding pattern and opiate activity.<br /> With the availability of monoclonal antibodies directed against IFN-?, it is possible to interpret results obtained from crude materials containing both IFN-? and IFNβ. The difficulties in studying in vitro and in vivo effects of 'type 1'. Interferons arise from the fact that both alpha and beta Interferons are produced in response to the same stimuli and also seem to act via the same receptor. These Interferon activities can only be distinguished from one another by use of specific neutralizing antibodies.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
F18
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Dalod et al. J Exp Med 2002;195:517
References 2:
Diebold et al. Nature 2003;424:324
References 3:
Joshi et al. J Interferon Cytokine Res 2006;26:739
MAP2a and 2b (270 kDa) being found mostly in dendrites, stabilize microtubules (shift the reaction kinetics in addition of new subunits and microtubule growth) and participate in determining the structure of different parts of vertebrate nerve cells.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
MT-01
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Mouse heavy chain of immunoglobulin (determined by immunodot). Avidity on IgM: 7 * 108 M-1. No crossreactivity with other animals. Available in unconjugated MON5057, HRP conjugated MON5057P
Mouse heavy chain of immunoglobulin (determined by immunodot). Avidity on IgM: 3.6 * 109 M-1. No crossreactivity with other animals. Available as unconjugated MON5057, PE conjugated MON 5057P, Biotin conjugated MON 5057B, FITC conjugated MON5057F
Antibody Isotype:
IgG1k
Monosan Range:
MONOSAN
Clone:
LO-MM-9
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin et al. Pergamon Press Oxford and NY 1982; 29:615-618
Mouse heavy chain of immunoglobulin (determined by immunodot). Avidity on IgM: 3.6 * 109 M-1. No crossreactivity with other animals. Available as unconjugated MON5057, PE conjugated MON 5057P, Biotin conjugated MON 5057B, FITC conjugated MON5057F
Antibody Isotype:
IgG1k
Monosan Range:
MONOSAN
Clone:
LO-MM-9
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin et al. Pergamon Press Oxford and NY 1982; 29:615-618
Mouse heavy chain of immunoglobulin (determined by immunodot). Avidity on IgM: 3.6 * 109 M-1. No crossreactivity with other animals. Available as unconjugated MON5057, PE conjugated MON 5057P, Biotin conjugated MON 5057B, FITC conjugated MON5057F
Antibody Isotype:
IgG1k
Monosan Range:
MONOSAN
Clone:
LO-MM-9
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin et al. Pergamon Press Oxford and NY 1982; 29:615-618
Mouse Gamma 3 Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG3: 2 * 1010 M-1. Crossreactivity with: Horse (+), swine (++) and dog (+/-). Available in unconjugated MON5056, FITC conjugated MON5056F, HRP conjugated MON5056H, Biotin conjugated MON5056B.
Mouse Gamma 3 Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG3: 2 * 1010 M-1. Crossreactivity with: Horse (+), swine (++) and dog (+/-). Available in unconjugated MON5056, FITC conjugated MON5056F, HRP conjugated MON5056H, Biotin conjugated MON5056B.
Mouse Gamma 3 Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG3: 2 * 1010 M-1. Crossreactivity with: Horse (+), swine (++) and dog (+/-). Available in unconjugated MON5056, FITC conjugated MON5056F, HRP conjugated MON5056H, Biotin conjugated MON5056B.
Mouse Gamma 3 Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG3: 2 * 1010 M-1. Crossreactivity with: Horse (+), swine (++) and dog (+/-). Available in unconjugated MON5056, FITC conjugated MON5056F, HRP conjugated MON5056H, Biotin conjugated MON5056B.
Mouse Gamma 2b Heavy Chain of Immunoglobulin (determined by immunodot). positively tested on BALB/c and C57BL/6 mouse sera. The specificity of every rat monoclonal antibody anti-mouse IgG subclasses is determined in a range of optimal concentrations. Increasing the first or the second antibody at an unnecessary too high concentration can induce possible cross-reactions with other mouse IgG subclasses. Avidity on IgG2b: 1 * 1010 M-1. No cross-reactivity with chicken, rabbit, goat, sheep, bovine, horse, dog, swine, baboon IgG and human IgG or IgM (ELISA test). Available unconjugated MON5055, FITC MON5055F, Biotin MON5055B, PE MON5055P
Mouse Gamma 2b Heavy Chain of Immunoglobulin (determined by immunodot). positively tested on BALB/c and C57BL/6 mouse sera. The specificity of every rat monoclonal antibody anti-mouse IgG subclasses is determined in a range of optimal concentrations. Increasing the first or the second antibody at an unnecessary too high concentration can induce possible cross-reactions with other mouse IgG subclasses. Avidity on IgG2b: 1 * 1010 M-1. No cross-reactivity with chicken, rabbit, goat, sheep, bovine, horse, dog, swine, baboon IgG and human IgG or IgM (ELISA test). Available unconjugated MON5055, FITC MON5055F, Biotin MON5055B, PE MON5055P
Mouse Gamma 2b Heavy Chain of Immunoglobulin (determined by immunodot). positively tested on BALB/c and C57BL/6 mouse sera. The specificity of every rat monoclonal antibody anti-mouse IgG subclasses is determined in a range of optimal concentrations. Increasing the first or the second antibody at an unnecessary too high concentration can induce possible cross-reactions with other mouse IgG subclasses. Avidity on IgG2b: 1 * 1010 M-1. No cross-reactivity with chicken, rabbit, goat, sheep, bovine, horse, dog, swine, baboon IgG and human IgG or IgM (ELISA test). Available unconjugated MON5055, FITC MON5055F, Biotin MON5055B, PE MON5055P
Mouse Gamma 2b Heavy Chain of Immunoglobulin (determined by immunodot). positively tested on BALB/c and C57BL/6 mouse sera. The specificity of every rat monoclonal antibody anti-mouse IgG subclasses is determined in a range of optimal concentrations. Increasing the first or the second antibody at an unnecessary too high concentration can induce possible cross-reactions with other mouse IgG subclasses. Avidity on IgG2b: 1 * 1010 M-1. No cross-reactivity with chicken, rabbit, goat, sheep, bovine, horse, dog, swine, baboon IgG and human IgG or IgM (ELISA test). Available unconjugated MON5055, FITC MON5055F, Biotin MON5055B, PE MON5055P
Mouse Gamma 2a Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG2a: 7 * 109 M-1. No crossreactivity with other animals. Available as unconjugated MON5054, HRP conjugated MON 5054P, Biotin conjugated MON 5054B
Mouse Gamma 2a Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG2a: 7 * 109 M-1. No crossreactivity with other animals. Available as unconjugated MON5054, HRP conjugated MON 5054P, Biotin conjugated MON 5054B
Mouse Gamma 2a Heavy Chain of Immunoglobulin (determined by immunodot). Avidity on IgG2a: 7 * 109 M-1. No crossreactivity with other animals. Available as unconjugated MON5054, HRP conjugated MON 5054P, Biotin conjugated MON 5054B
Mouse gamma 1 heavy chain of immunoglobulin (determined by immunodot). No crossreactivity with other animals. Available as unconjugated MON5053, FITC conjugated MON 5053F, HRP conjugated Cat.no. MON 5053P, Biotin conjugated MON 5053B.
Antibody Isotype:
IgGk
Monosan Range:
MONOSAN
Clone:
LO-MG1-2
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin, et al., J.Immunology Meth. 1984;71, 9-16
References 2:
Querinjean, et al.,. Eur.J.Biochem. 1972: 31:354-359
Mouse gamma 1 heavy chain of immunoglobulin (determined by immunodot). No crossreactivity with other animals. Available as unconjugated MON5053, FITC conjugated MON 5053F, HRP conjugated Cat.no. MON 5053P, Biotin conjugated MON 5053B.
Antibody Isotype:
IgGk
Monosan Range:
MONOSAN
Clone:
LO-MG1-2
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin, et al., J.Immunology Meth. 1984;71, 9-16
References 2:
Querinjean, et al.,. Eur.J.Biochem. 1972: 31:354-359
Mouse gamma 1 heavy chain of immunoglobulin (determined by immunodot). No crossreactivity with other animals. Available as unconjugated MON5053, FITC conjugated MON 5053F, HRP conjugated Cat.no. MON 5053P, Biotin conjugated MON 5053B.
Antibody Isotype:
IgGk
Monosan Range:
MONOSAN
Clone:
LO-MG1-2
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin, et al., J.Immunology Meth. 1984;71, 9-16
References 2:
Querinjean, et al.,. Eur.J.Biochem. 1972: 31:354-359
Mouse gamma 1 heavy chain of immunoglobulin (determined by immunodot). No crossreactivity with other animals. Available as unconjugated MON5053, FITC conjugated MON 5053F, HRP conjugated Cat.no. MON 5053P, Biotin conjugated MON 5053B.
Antibody Isotype:
IgGk
Monosan Range:
MONOSAN
Clone:
LO-MG1-2
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide
Storage:
-20°C
References 1:
Bazin, et al., J.Immunology Meth. 1984;71, 9-16
References 2:
Querinjean, et al.,. Eur.J.Biochem. 1972: 31:354-359
Mouse epsilon heavy chain of immunoglobulin (determined by immunodot). Useful as second antibody in immunoassays, such as ELISA's (good binding on plastic) and immunohistochemistry on frozen sections. Available in unconjugated MON5051, FITC conjugated MON5051F, HRP conjugated MON5050P and Biotin conjugated MON5051B.
Mouse epsilon heavy chain of immunoglobulin (determined by immunodot). Useful as second antibody in immunoassays, such as ELISA's (good binding on plastic) and immunohistochemistry on frozen sections. Available in unconjugated MON5051, FITC conjugated MON5051F, HRP conjugated MON5050P and Biotin conjugated MON5051B.
Mouse epsilon heavy chain of immunoglobulin (determined by immunodot). Useful as second antibody in immunoassays, such as ELISA's (good binding on plastic) and immunohistochemistry on frozen sections. Available in unconjugated MON5051, FITC conjugated MON5051F, HRP conjugated MON5050P and Biotin conjugated MON5051B.
Mouse epsilon heavy chain of immunoglobulin (determined by immunodot). Useful as second antibody in immunoassays, such as ELISA's (good binding on plastic) and immunohistochemistry on frozen sections. Available in unconjugated MON5051, FITC conjugated MON5051F, HRP conjugated MON5050P and Biotin conjugated MON5051B.
Mouse alpha heavy chain of immunoglobulin (determined by immunodot). Avidity on IgA: 2.9 * 109 M-1. Available as unconjugated MON5050, Biotin conjugated MON5050B, Peroxidase conjugated MON5050P.
Antibody Isotype:
IgMk
Monosan Range:
MONOSAN
Clone:
LO-MA-7
Concentration:
1 mg/ml
Storage buffer:
PBS with 0.1% sodium azide 50% glycerol
Storage:
-20°C
References 1:
Bazin et al. Pergamon Press Oxford and NY 1982; 29:615-618
Mouse alpha heavy chain of immunoglobulin (determined by immunodot). Avidity on IgA: 4.8 * 109 M-1. Useful as second antibody in immunoassays, such as ELISA's (good binding on plastic) and immunohistochemistry on frozen sections. Available in unconjugated MON5050, HRP conjugated MON5050P, Biotin conjugated MON5050B
Mouse alpha heavy chain of immunoglobulin (determined by immunodot). Avidity on IgA: 4.8 * 109 M-1. Useful as second antibody in immunoassays, such as ELISA's (good binding on plastic) and immunohistochemistry on frozen sections. Available in unconjugated MON5050, HRP conjugated MON5050P, Biotin conjugated MON5050B
The monoclonal antibody 7C2 can be used during various purification steps of IgY. The yolk of eggs laid by immunized chickens has been recognized as an excellent source of polyclonal antibodies (pAb). Specific antibodies produced in chickens offer several important advantages over producing antibodies in other mammals. Because a single egg contains as much antibody as an average bleed from a rabbit, this simple, non-invasive approach presents an appealing alternative to conventional pAb production methods. Purification of chicken egg yolk immunoglobulin Y (IgY), the 150 kDa IgG homolog, does not require animal bleeding. In addition, the eggs from immunized chickens provide a continual, daily source of pAb, and this convenient approach offers greater compatibility with animal protection regulations. Due to the phylogenetic distance between birds and mammals, there is greater potential of producing a higher percentage of specific antibody against mammalian antigens when using chickens. Highly conserved mammalian proteins sometimes fail to illicit a humoral response in animals, such as rabbits, that are traditionally used for generating pAb. Non-specific binding and need for cross-species immunoabsorptions is eliminated since chicken IgY does not cross-react with mammalian IgG and does not bind bacterial or mammalian Fc receptors. There are well defined structural differences of IgY-type immunoglobulins and the IgG of mammals. That includes the molar mass of the heavy chains of the immunoglobulins. The IgY-type immunoglobulins are much less flexible than IgG. Also, the structures of the Fc part of the immunoglobulin isotypes IgY and IgG are different. The antibody 7C2 is cross reactive for duck IgY.
The monoclonal antibody 7C2 can be used during various purification steps of IgY. The yolk of eggs laid by immunized chickens has been recognized as an excellent source of polyclonal antibodies (pAb). Specific antibodies produced in chickens offer several important advantages over producing antibodies in other mammals. Because a single egg contains as much antibody as an average bleed from a rabbit, this simple, non-invasive approach presents an appealing alternative to conventional pAb production methods. Purification of chicken egg yolk immunoglobulin Y (IgY), the 150 kDa IgG homolog, does not require animal bleeding. In addition, the eggs from immunized chickens provide a continual, daily source of pAb, and this convenient approach offers greater compatibility with animal protection regulations. Due to the phylogenetic distance between birds and mammals, there is greater potential of producing a higher percentage of specific antibody against mammalian antigens when using chickens. Highly conserved mammalian proteins sometimes fail to illicit a humoral response in animals, such as rabbits, that are traditionally used for generating pAb. Non-specific binding and need for cross-species immunoabsorptions is eliminated since chicken IgY does not cross-react with mammalian IgG and does not bind bacterial or mammalian Fc receptors. There are well defined structural differences of IgY-type immunoglobulins and the IgG of mammals. That includes the molar mass of the heavy chains of the immunoglobulins. The IgY-type immunoglobulins are much less flexible than IgG. Also, the structures of the Fc part of the immunoglobulin isotypes IgY and IgG are different. The 7C2 antibody is cross reactive for duck
The antibody recognizes an epitope that is exposed by protein-biotin and nucleic acid-biotin conjugates. The antibody can be used in molecular biological tests and immunohistology. The antibody permits the formation of antibody-biotin-streptavidin-enzyme complexes thus enhancing the sensitivity of the detection system.
Alpha-fetoprotein (AFP) is present in fetal plasma, and it binds e.g. copper, nickel, and bilirubin. Measuring of alpha-fetoprotein level in amniotic fluid can reveal severe fetal defects. In adults, elevated AFP concentrations in the plasma can indicate hepatocellular carcinoma or teratoblastoma. In some individuals, hereditary persistance of alpha-fetoprotein can be observed without any obvious pathology.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
AFP-11
Concentration:
1 mg/ml
Format:
Purified by protein-A affinity chromatography.
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Alpha-fetoprotein (AFP) is present in fetal plasma, and it binds e.g. copper, nickel, and bilirubin. Measuring of alpha-fetoprotein level in amniotic fluid can reveal severe fetal defects. In adults, elevated AFP concentrations in the plasma can indicate hepatocellular carcinoma or teratoblastoma. In some individuals, hereditary persistance of alpha-fetoprotein can be observed without any obvious pathology.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
AFP-01
Concentration:
1 mg/ml
Format:
Purified by protein-A affinity chromatography.
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Human serum albumin (65-67 kDa) is the most abundant protein in human blood plasma (produced in the liver). It has a serum half-life of approximately 20 days.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
AL-01
Conjugate:
Biotin
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Human serum albumin (65-67 kDa) is the most abundant protein in human blood plasma (produced in the liver). It has a serum half-life of approximately 20 days.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
AL-01
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
MON5023 reacts only with mature insulin and has no reactivity with the C-Peptide and insulin chains. Insulin and glucagon are pancreatic endocrine hormones secreted by islet cells within the pancreas. The stimulus for insulin secretion is a HIGH blood glucose. Deficiency of insulin results in diabetes mellitus, one of the leading causes of morbidity and mortality in the general population.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
IN-05
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The antibody reacts with human native and recombinant IL-6 as assessed by ELISA. The antibody inhibits the biological activity of human native and recombinant IL-6 as determined with the B9 cell bioassay. The antibody cross reacts with rhesus and cynomolgus natural IL6.
From every molecule of proinsulin, one molecule of insulin plus one molecule of C-peptide are produced. C-peptide is released into the blood stream in equal amounts to insulin.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
C-PEP-01
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
The antibody neutralizes mouse interleukin 6 (IL-6). It does not cross-react with LPS (the component of the bacterial cell wall which is considered responsible for the toxicity of gram-negative bacteria) and TNFalpha nor does it block human or rat IL-6 in proliferation assays using the KD83 cell line. MP5-32C11 inhibits mouse IL-6-induced proliferation of the NFS60 myelomonocytic cell line and KD83 plasmacytoma. IL-6 is a pluripotent 20-22 kDa cytokine which plays a role in the pathophysiology of severe infection and regulates the immune response, acute phase reaction and haematopoiesis. IL-6 plays a critical role in Bcell differentiation to plasma cells and is a potent growth factor for plasmacytoma and myeloma. Continuous IL-6 gene expression makes an essential contribution to a multistep oncogenesis of plasma cell neoplasia. IL-6 is a very useful culture supplement for the generation of a high number of antibodyproducing hybridomas.
Transferrin is a monomeric glycoprotein of approximately 77 kDa, which serves as an iron-transporter. In normal plasma, transferrin has a concentration of 25-50 µmol / liter, and is usually about one-third saturated with iron, thus providing a large buffering capacity in case of an acute increase in plasma iron levels. Cells take up transferrin-iron complexes (holotransferrin) using transferrin receptor dimers. Upon binding of holotransferrin, the receptor is internalized by clathrin-mediated endocytosis. Acidification of endosomes by vesicular membrane proton pumps leads to dissociation of iron ions, whereas transferrin (apotransferrin) remains associated with its receptor (CD71) and recycles to the cell surface, where apotransferrin is released upon exposure to normal pH. Internalization of labeled transferrin thus represents an usefull approach to study endocytosis. Serum concentration rises in iron deficiency and pregnancy and falls in iron overload, infection and inflammatory conditions. Iron/transferrin complex is essential in haemoglobin synthesis and for certain types of cell division.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
HTF-14
Concentration:
1 mg/ml
Format:
Purified by protein-A affinity chromatography.
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Storage:
2-8°C
References 1:
Bartek J., et al., Immunol. Lett. 1982; 7: 231.
References 2:
Bartek, J., et al., Br. J. Haematol 1985; 59: 435-441.
References 3:
Nováková, M., et al., Cell Motil Cytoskeleton 1996;33(1):38-51
Mab MH25-1 reacts specifically with human IgE e-chains; De-1 epitope on Fc fragment. No cross reaction with other immunoglobulins. Positive control: Polypus nose
MH14-1 recognizes IgA heavy chains; epitope present on IgA<sub>1</sub> and IgA<sub>2</sub>. No cross reactivity with IgG or IgM (tested with ELISA).Suitable for lymphoma phenotyping. Recommended for positive control: Duodenum
The monoclonal antibody 52B83 reacts with tumor necrosis factor alpha (TNF-alpha). TNF-alpha is a homotrimeric 17 kDa protein, that interacts with either one of the two types of TNF-receptors, termed I and II, leading to receptor cross-linking and signal transduction. The receptors differ strongly in their intra-cellular signaling pathways.<br /> TNF-alpha was originally described as a highly cytotoxic cytokine for tumor cells, it causes tumor necrosis in vivo and shows cytolytic activity against tumor cells in vitro. Furthermore,- TNF-alpha is found to be a central mediator in many inflammatory and immunological processes. It can be induced by various products of micro-organisms and by various cytokines leading to expression of a wide variety of- cytokines. The pro-inflammatory properties of- TNF-alpha play a central role in several auto-immune diseases such as rheumatoid arthritis and inhibition by neutralizing molecules have been shown to be beneficial in patients.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
52B83
Concentration:
100 ug/ ml
Storage buffer:
PBS with 0.1% BSA and 0.02% sodium azide
Storage:
2-8°C
References 1:
Bradding; P et al. Am J Respir Cell Mol Biol 1994; 10: 471
References 2:
Bradding, P et al: Clin Exp Allergy 1995, 25: 406
References 3:
Gerspach; J et al. Microsc Res Tech 2000; 50: 243
References 4:
Laan van der N et al. Arch Dermatol Res 2001; 293: 226
The antibody will stain lambda chain-containing human cells. In ELISA the antibody will react with free lambda chains and also with lambda chains attached to heavy chains. Membrane bound Ig containing lambda light chains will also be recognized as well as cytoplasmic Ig from bone marrow cells containing lambda chains. All cells reacting with conventional anti-lambda will also react with antibody. No crossreactivity with kappa chains detected.
The antibody reacts with free kappa chains, but not with kappa chains attached to heavy chains. No reaction is observed with Ig kappa on mem¬branes of lymphocytes. No crossreactivity with lambda chains. In Ouchterlony immunodiffusion no reaction with Bence Jones kappa is observed.
Daxx is an apoptosis-modulating death domain-associated protein with functions in transcriptional regulation. Daxx functions both in cytoplasm, where it interacts with Fas, and in nucleus (residing in PML oncogenic domains), where it is involved in SUMO-dependent regulation of transcription and subnuclear compartmentalization. Daxx senzitizes the cells to apoptosis, but on the other hand, this protein may also serve in preventing apoptosis in the early embryo. Even regarding the transcription, Daxx can serve both as a corepressor and a coactivator. During ischemic stress, Daxx translocates from the nucleus to the cytoplasm, where in regulates sodium hydrogen exchanger isoform 1 (NHE1).
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
DAXX-01
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Mouse anti Giardia lamblia antibody, clone 6231 recognizes Giardia lamblia, a flagellated protozoan parasite which infects the small intestines of humans and a variety of other mammalian hosts. Infection by G. lamblia results in giardiasis; a type of gastroenteritis that causes severe diarrhea and abdominal cramps. It has been suggested that chronic giardiasis can result in long-term growth retardation (Simsek. et al. 2004).
Mouse anti Human CD81 antibody, clone 1D6 recognizes human CD81, a 26 kDa cell surface antigen also known as TAPA-1, and a member of the tetraspanin family. CD81 is widely expressed on human leucocytes and appears to be involved in a variety of cellular leucocytes including activation, proliferation and differentiation.Mouse anti Human CD81 antibody, clone 1D6 is a potent CD81 reagent, induces homotypic adhesion and has powerful anti-proliferative effects.
Mouse anti Human CD226 antibody, clone DX11 recognizes human CD226, a ~65 kDa glycoprotein, also known as DNAM1 (DNAX accessory molecule-1). CD226 is broadly expressed on T-cells, NK cells, platelets, monocytes and a subset of B cells. CD226 is also expressed by a subset of CD3 positive thymocytes.Mouse anti Human CD226 antibody, clone DX11 is reported to inhibit T- and NK cell mediated cytotoxicity against tumor cell targets and to block TNF alpha and IFN gamma secretion by alloantigen-specific T-cells (Kojima et al. 2003).
Mouse anti Human CD226 antibody, clone DX11 recognizes human CD226, a ~65 kDa glycoprotein, also known as DNAM1 (DNAX accessory molecule-1). CD226 is broadly expressed on T-cells, NK cells, platelets, monocytes and a subset of B cells. CD226 is also expressed by a subset of CD3 positive thymocytes.Mouse anti Human CD226 antibody, clone DX11 is reported to inhibit T- and NK cell mediated cytotoxicity against tumor cell targets and to block TNF alpha and IFN gamma secretion by alloantigen-specific T-cells (Kojima et al. 2003).
Mouse anti Human CD226 antibody, clone DX11 recognizes human CD226, a ~65 kDa glycoprotein, also known as DNAM1 (DNAX accessory molecule-1). CD226 is broadly expressed on T-cells, NK cells, platelets, monocytes and a subset of B cells. CD226 is also expressed by a subset of CD3 positive thymocytes.Mouse anti Human CD226 antibody, clone DX11 is reported to inhibit T- and NK cell mediated cytotoxicity against tumor cell targets and to block TNF alpha and IFN gamma secretion by alloantigen-specific T-cells (Kojima et al. 2003).
Mouse anti Human CD226 antibody, clone DX11 recognizes human CD226, a ~65 kDa glycoprotein, also known as DNAM1 (DNAX accessory molecule-1). CD226 is broadly expressed on T-cells, NK cells, platelets, monocytes and a subset of B cells. CD226 is also expressed by a subset of CD3 positive thymocytes.Mouse anti Human CD226 antibody, clone DX11 is reported to inhibit T- and NK cell mediated cytotoxicity against tumor cell targets and to block TNF alpha and IFN gamma secretion by alloantigen-specific T-cells (Kojima et al. 2003).
Mouse anti Human CD101 antibody, clone BB27 recognizes human CD101, also known as Immunoglobulin superfamily member 2 (IgSF2), . Cell surface glycoprotein V7, Glu-Trp-Ile EWI motif-containing protein 101 or EWI-101. CD101 is a 1021 amino acid, includinng a 20 amino acid signal peptide, ~140 kDa single pass type I homodimeric cell surface glycoprotein expressed primarily by monocytes, granulocytes, dendritic cells and activated T lymphocytes.CD101 plays a major role in the activation of T cells by skin dendritic cells. Mouse anti Human CD101 antibody, clone BB27 has been reported to inhibit allogeneic T-cell responses (Bagot et al. 1997).
Mouse anti Human CD101 antibody, clone BB27 recognizes human CD101, also known as Immunoglobulin superfamily member 2 (IgSF2), . Cell surface glycoprotein V7, Glu-Trp-Ile EWI motif-containing protein 101 or EWI-101. CD101 is a 1021 amino acid, includinng a 20 amino acid signal peptide, ~140 kDa single pass type I homodimeric cell surface glycoprotein expressed primarily by monocytes, granulocytes, dendritic cells and activated T lymphocytes.CD101 plays a major role in the activation of T cells by skin dendritic cells. Mouse anti Human CD101 antibody, clone BB27 has been reported to inhibit allogeneic T-cell responses (Bagot et al. 1997).
Mouse anti Human CD101 antibody, clone BB27 recognizes human CD101, also known as Immunoglobulin superfamily member 2 (IgSF2), . Cell surface glycoprotein V7, Glu-Trp-Ile EWI motif-containing protein 101 or EWI-101. CD101 is a 1021 amino acid, includinng a 20 amino acid signal peptide, ~140 kDa single pass type I homodimeric cell surface glycoprotein expressed primarily by monocytes, granulocytes, dendritic cells and activated T lymphocytes.CD101 plays a major role in the activation of T cells by skin dendritic cells. Mouse anti Human CD101 antibody, clone BB27 has been reported to inhibit allogeneic T-cell responses (Bagot et al. 1997).
Rat anti Mouse CD79b antibody, clone AT107-2 recognizes a cytoplasmic region of mouse B-cell antigen receptor complex-associated protein beta chain, also known as B-cell-specific glycoprotein B29, Ig-beta, or Immunoglobulin-associated B29 protein. CD79b is a 228 amino acid, including a 26 signal peptide ~40 kDa type I single pass transmembrane glycoprotein. CD79b is expressed by B lymphocytes and associates with CD79a to form a heterodimer, non-covalently linked to surface immunoglobulin, forming the B-cell receptor (BCR) complex. Rat anti Mouse CD79b antibody, clone AT107-2 also recognizes a homologous region of human CD79b.
Rat anti Mouse CD79b antibody, clone AT107-2 recognizes a cytoplasmic region of mouse B-cell antigen receptor complex-associated protein beta chain, also known as B-cell-specific glycoprotein B29, Ig-beta, or Immunoglobulin-associated B29 protein. CD79b is a 228 amino acid, including a 26 signal peptide ~40 kDa type I single pass transmembrane glycoprotein. CD79b is expressed by B lymphocytes and associates with CD79a to form a heterodimer, non-covalently linked to surface immunoglobulin, forming the B-cell receptor (BCR) complex. Rat anti Mouse CD79b antibody, clone AT107-2 also recognizes a homologous region of human CD79b.
CD206 (macrophage mannose receptor, MMR), also known as mannose receptor C1 (MRC1), is a type I transmembrane glycoprotein serving as pattern recognition receptor for carbogydrate groups on the surface of bacteria, fungi and other pathogens. Expressed mainly on tissue macrophages and dendritic cells, CD206 mediates endocytosis of these pathogens and presentation of their antigens to the adaptive immune system. CD206 can also be detected in a soluble form in human plasma and is elevated in patients with acute sepsis.
Rat anti Human CD195 antibody, clone HEK/1/85a recognizes the human CD195 cell surface antigen, a 45 kDa glycoprotein also known as CCR5.CD195 acts as a receptor for a number of chemokines including RANTES and eotaxin and serves as a co-receptor for the entry of HIV into cells. CD195 is expressed by a subset of T lymphocytes and by monocytes.
Rat anti Human CD195 antibody, clone HEK/1/85a recognizes the human CD195 cell surface antigen, a 45 kDa glycoprotein also known as CCR5.CD195 acts as a receptor for a number of chemokines including RANTES and eotaxin and serves as a co-receptor for the entry of HIV into cells. CD195 is expressed by a subset of T lymphocytes and by monocytes.
Rat anti Human CD195 antibody, clone HEK/1/85a recognizes the human CD195 cell surface antigen, a 45 kDa glycoprotein also known as CCR5.CD195 acts as a receptor for a number of chemokines including RANTES and eotaxin and serves as a co-receptor for the entry of HIV into cells. CD195 is expressed by a subset of T lymphocytes and by monocytes.
Mouse anti Human CD146 antibody, clone OJ79c recognizes human Cell surface glycoprotein MUC18, also known as CD146, Cell surface glycoprotein P1H12, Melanoma cell adhesion molecule (MCAM) or S-endo 1 endothelial-associated antigen. CD146 is a 646 amino acid single pass type 1 transmembrane glycoprotein with a calculated molecular mass of ~72 kDa. However due to extensive N-linked glycosylation CD146 migrates in polyacrylamide gels with an apparent molecular mass of ~118 kDa. CD146 is a member of the immunoglobulin superfamily bearing 2 V-type Ig-like and 3 C-type Ig-like domains. CD146 is expressed by all endothelial cells and by melanoma cells and appears to act as an adhesion molecule (UniProt: P43121). Expression in melanoma may be linked to tumor progression (Lehmann et al. 1989).Mouse anti Human CD146 antibody, clone OJ79c is highly expressed on pericytes and has been utilized for the identification of perivascular mesenchymal precursor cells from cardiac muscle using flow cytometry (Chen et al. 2014).
Mouse anti Human CD146 antibody, clone OJ79c recognizes human Cell surface glycoprotein MUC18, also known as CD146, Cell surface glycoprotein P1H12, Melanoma cell adhesion molecule (MCAM) or S-endo 1 endothelial-associated antigen. CD146 is a 646 amino acid single pass type 1 transmembrane glycoprotein with a calculated molecular mass of ~72 kDa. However due to extensive N-linked glycosylation CD146 migrates in polyacrylamide gels with an apparent molecular mass of ~118 kDa. CD146 is a member of the immunoglobulin superfamily bearing 2 V-type Ig-like and 3 C-type Ig-like domains. CD146 is expressed by all endothelial cells and by melanoma cells and appears to act as an adhesion molecule (UniProt: P43121). Expression in melanoma may be linked to tumor progression (Lehmann et al. 1989).Mouse anti Human CD146 antibody, clone OJ79c is highly expressed on pericytes and has been utilized for the identification of perivascular mesenchymal precursor cells from cardiac muscle using flow cytometry (Chen et al. 2014).
CD229 (Ly9) is a cell surface receptor of the CD150 family, which includes also e.g. CD48 and CD224. Receptors of this family regulate cytokine production and cytotoxicity of lymphocytes and NK cells. High levels of CD229 are found on T and B cells, where its expression increases during their maturation. It is absent on granulocytes, bone marrow-derived dendritic cells, platelets and erythrocytes. CD229 has been also reported on mouse monocytes and NK cells. CD229 interacts homophilically through its N-terminal domain and localizes to the contact site between T cells and antigen presenting B cells during antigen-dependent immune synapse formation.
Mouse anti Human CD229 antibody, clone HLy9.1.25 recognizes the human cell surface antigen CD229 also known as T-lymphocyte surface antigen Ly-9 or SLAM family member 3. CD229 is a 608 amino acid single pass type I transmembrane glycoprotein of ~120 kDa as evaluated by immunoprecipitation of cells transfected with the full length human CD229 cDNA. However immunoprecipitation of CD229 from Daudi cell lysates with dlone HLy9.1.25 yields bands of 120 kDa corresponding to the full length CD229 and a ~100 kDa band attributed to an alternatively spliced isoform lacking the fourth Ig-like domain (de la Fuente et al. 2001).Human CD299 is expressed on thymocytes, T-cells and B-cells (Del Valle et al. 2003). CD229 has also been described as a tumor associated antigen in chronic lymphocytic leukemia (Bund et al. 2006) and has been implicated in the development of spontaneous autoantibody production to nuclear antigens in mice and is potentially a target for the treatment of autoimmunity (de Salort et al. 2013).
Mouse anti Human CD229 antibody, clone HLy9.1.25 recognizes the human cell surface antigen CD229 also known as T-lymphocyte surface antigen Ly-9 or SLAM family member 3. CD229 is a 608 amino acid single pass type I transmembrane glycoprotein of ~120 kDa as evaluated by immunoprecipitation of cells transfected with the full length human CD229 cDNA. However immunoprecipitation of CD229 from Daudi cell lysates with dlone HLy9.1.25 yields bands of 120 kDa corresponding to the full length CD229 and a ~100 kDa band attributed to an alternatively spliced isoform lacking the fourth Ig-like domain (de la Fuente et al. 2001).Human CD299 is expressed on thymocytes, T-cells and B-cells (Del Valle et al. 2003). CD229 has also been described as a tumor associated antigen in chronic lymphocytic leukemia (Bund et al. 2006) and has been implicated in the development of spontaneous autoantibody production to nuclear antigens in mice and is potentially a target for the treatment of autoimmunity (de Salort et al. 2013).
Mouse anti Human CD88 antibody, clone P12/1 recognizes the C5a receptor (C5aR) also known as CD88 or C5a anaphylatoxin chemotactic receptor 1. CD88 is predominantly expressed on cells of the myeloid lineage. When C5aR is preincubated with C5a, Mouse anti Human CD88 antibody, clone P12/1 does not bind to the receptor, as the binding site of P12/1 is located in the C5a binding region (Werfel et al. 1996 and Weinman et al. 2003)
Mouse anti Human CD88 antibody, clone P12/1 recognizes the C5a receptor (C5aR) also known as CD88 or C5a anaphylatoxin chemotactic receptor 1. CD88 is predominantly expressed on cells of the myeloid lineage. When C5aR is preincubated with C5a, Mouse anti Human CD88 antibody, clone P12/1 does not bind to the receptor, as the binding site of P12/1 is located in the C5a binding region (Werfel et al. 1996 and Weinman et al. 2003)
Mouse anti Human CD238 antibody, clone BRIC203 recognizes human CD238, also known as the kell blood group antigen Kpbc. The kell blood group antigens are carried on the ~93 kDa erythrocyte membrane glycoprotein.In hemagglutination tests, Mouse anti Human CD238 antibody, clone BRIC203 reacts with Kp (a-b+) but not Kp (a+b-). In addition Mouse anti Human CD238 antibody, clone BRIC203 will agglutinate Kp (a+b-c+) and Kp (a-b-c+) erythrocytes but not react with Ko erythrocytes. Weaker staining in flow cytometry is observed on erythroblasts derived from CD34+ hematopoietic progenitor cells, compared to Kp (a-b+) red blood cells.
Mouse anti Human CD175 antibody, clone BRIC111 recognizes Tn antigen on glycophorin A and glycophorin B in human erythrocytes. Tn is a cryptantigen which was designated CD175 at the 7th Leucocyte Typing Workshop. Tn antigen is not expressed on normal haemopoietic cells but exposure of the Tn is associated with polyagglutination.
Mouse anti Human CD173 antibody, clone BRIC231 recognizes human type 2 H blood group antigen, also known as CD173. Active H substances in man, are expressed by many cells and tissues and also by erythrocytes.
Mouse anti Human CD236 antibody, clone BGRL100 recognizes a 17 amino acid residue within the cytoplasmic domain of glycophorin C (GPC), which has recently been designated CD236.CD236 is an integral membrane protein which carries the Gerbich blood group antigens. Expression of CD236 is not restricted to erythroid cells, but is broadly distributed in both erythroid and non-erythroid tissues.
Mouse anti Human CD239 antibody, clone BRIC221 recognizes human CD239, also known as Lutheran antigen or basal cell aghesion molecule.CD239 is a 597 amino acid, ~85 kDa single pass type I membrane glycoprotein. Clone BRIC221 recognizes a monomorphic determinant expresses on both the 85 and 78 kDa Lutheran (Lu) glycoforms (El Nemer et al. 1998). BRIC 221 recognizes an epitope in the fourth extracellular domain of Lu glycoprotein (Parsons et al. 1997). Lutheran glycoprotein is a member of the immunoglobulin superfamily and was designated CD239 (B-CAM) at the 7th leucocyte typing workshop. CD239 is expressed by erythrocytes in the peripheral blood.
Mouse anti Human CD154 antibody, clone MK13A4 recognizes the human CD154 cell surface antigen, a 32 kDa glycoprotein also known as CD40 ligand. CD154 is expressed on activated T lymphocytes, predominantly CD4+ve cells and also on some basophils and mast cells.Mouse anti Human CD154 antibody, clone MK13A4 has been reported to block binding of CD40 ligand to its receptor CD40.
Mouse anti Human CD91 antibody, clone A2Mr alpha-2 recognizes human CD91, also known as Prolow-density lipoprotein receptor-related protein 1, Alpha-2-macroglobulin receptor or apolipoprotein E receptor. CD91 is a 4525 amino acid protein post translationally cleaved into 3 subunits, a 85 kDa type I transmembrane carboxyl chain (LRP85) non-covalently bound to a 515 kDa extracellular N-terminal subunit (LRP515)containing multiple EGF-like and LDL-receptor Class A and Class B domains. Additionally, there is an intracellular domain (LRPICD) which can be cleaved from the transmambrane domain by gamma secretase (May et al. 2004). Clone A2Mr alpha-2 detects an epitope within the LRP515 chain.CD91 is a multifunctional protein involved in processes inluding the phagocytosis and endocytosis of apoptotic cells (Nilsson et al. 2012), clearance of activated serum alpha-2-macroglobulin (Kristensen et al. 1990), modulation of the inflammatory response (Staudt et al. 2013) and acts as a receptor for Pseudomonas aeruginosa exotoxin A (Kounnas et al. 1992).Mouse anti Human CD91, clone A2Mr alpha-2 has been used extensively for the detection of CD91 by flow cytometry and immunohistochemistry
Mouse anti Human CD91 antibody, clone A2Mr alpha-2 recognizes human CD91, also known as Prolow-density lipoprotein receptor-related protein 1, Alpha-2-macroglobulin receptor or apolipoprotein E receptor. CD91 is a 4525 amino acid protein post translationally cleaved into 3 subunits, a 85 kDa type I transmembrane carboxyl chain (LRP85) non-covalently bound to a 515 kDa extracellular N-terminal subunit (LRP515)containing multiple EGF-like and LDL-receptor Class A and Class B domains. Additionally, there is an intracellular domain (LRPICD) which can be cleaved from the transmambrane domain by gamma secretase (May et al. 2004). Clone A2Mr alpha-2 detects an epitope within the LRP515 chain.CD91 is a multifunctional protein involved in processes inluding the phagocytosis and endocytosis of apoptotic cells (Nilsson et al. 2012), clearance of activated serum alpha-2-macroglobulin (Kristensen et al. 1990), modulation of the inflammatory response (Staudt et al. 2013) and acts as a receptor for Pseudomonas aeruginosa exotoxin A (Kounnas et al. 1992).Mouse anti Human CD91, clone A2Mr alpha-2 has been used extensively for the detection of CD91 by flow cytometry and immunohistochemistry
Mouse anti Human CD200 antibody, clone OX-104 recognizes the human CD200 cell surface antigen, also known as OX2.CD200 is expressed by a subset of B lymphocytes, some endothelial cells and by neurons. Studies have suggested that the CD200-CD200 ligand system is of importance in the control of macrophage and granulocyte activation.
Mouse anti Human CD200 antibody, clone OX-104 recognizes the human CD200 cell surface antigen, also known as OX2.CD200 is expressed by a subset of B lymphocytes, some endothelial cells and by neurons. Studies have suggested that the CD200-CD200 ligand system is of importance in the control of macrophage and granulocyte activation.
Mouse anti Human CD112 antibody, clone R2-525 recognizes CD112, also known as Poliovirus Receptor Related 2, PRR2 or Nectin-2, a surface molecule originally reported to be homologous to CD155, the poliovirus receptor (Lopez et al. 1998). PRR2 expression is restricted to the myelo-monocytic and megakaryocytic lineages. PRR2 expression has also been detected on haematopoeitic progenitors expressing CD34.
Mouse anti Human CD94 antibody, clone DX22 recognizes human Natural killer cells antigen CD94, also known as KLRD1, Killer cell lectin-like receptor, subfamily D, member 1, CD94, KP43 and NK cell receptor. CD94 is a 179 amino acid single pass type II transmembrane glycoprotein with a predicted molecular mass of ~20.5 kDa. There are 3 isoforms of CD94 derived by alternative splicing.CD94 is expressed on natural killer (NK) cells and a subset of T lymphocytes.CD94 is found to associate with NKG2 to form a heterodimer, involved in the inhibition of cell mediated cytotoxicity against cells bearing appropriate MHC class I allotypes.Mouse anti Human CD94 antibody, clone DX22 is reported to inhibit the binding of CD94 to HLA-E (Braud et al. 1998) and HLA-G (Söderström. et al. 1997).
Mouse anti Human CD89 antibody, clone MIP8a recognizes the human CD89 cell surface antigen, a 50-75 kDa cell surface glycoprotein that is also known as the IgA receptor (Fc alpha R).CD89 is expressed by peripheral blood monocytes and neutrophils.MIP8a blocks binding of IgA to the Fc alpha R, and also inhibits neutrophil phagocytosis of IgA complexes.
Mouse anti Human CD89 antibody, clone MIP8a recognizes the human CD89 cell surface antigen, a 50-75 kDa cell surface glycoprotein that is also known as the IgA receptor (Fc alpha R).CD89 is expressed by peripheral blood monocytes and neutrophils.MIP8a blocks binding of IgA to the Fc alpha R, and also inhibits neutrophil phagocytosis of IgA complexes.
Mouse anti Human CD227 antibody, clone C595 (NCRC48) recognizes CD227, also known as mucin 1 which is a breast cancer associated mucin encoded by the Muc-1 gene. Mucins are a family of high molecular weight, heavily glycosylated proteins (glycoconjugates) produced by many epithelial tissues in vertebrates. CD227 is expressed on most secretory epithelium, including mammary gland and some hematopoietic cells. This protein is overexpressed abundantly in >90% breast carcinomas and metastases.Mouse anti Human CD227 antibody, clone C595 recognizes the peptide epitope ARG-PRO-ALA-PRO within the protein core of the mucin.
Mouse anti Human CD227 antibody, clone C595 (NCRC48) recognizes CD227, also known as mucin 1 which is a breast cancer associated mucin encoded by the Muc-1 gene. Mucins are a family of high molecular weight, heavily glycosylated proteins (glycoconjugates) produced by many epithelial tissues in vertebrates. CD227 is expressed on most secretory epithelium, including mammary gland and some hematopoietic cells. This protein is overexpressed abundantly in >90% breast carcinomas and metastases.Mouse anti Human CD227 antibody, clone C595 recognizes the peptide epitope ARG-PRO-ALA-PRO within the protein core of the mucin.
Mouse anti Human CD152 antibody, clone BNI3 recognizes human CD152, also known as CTLA-4 (cytotoxic T-lymphocyte-associated antigen 4), an inhibitory receptor and negative regulator of T-cell responses. CD152 is a single pass type 1 transmembrane protein belonging to the immunoglobulin superfamily containing a single Ig-v-like domain in the extracellular region.CD152 along with CD28 binds to the co-stimulatory molecules CD80 and CD86 (Azuma et al. 1993). Mouse anti human CD152 antibody, clone BNI3 is able to block ligand binding on the Raji B-cell line (Steiner et al. 2001) and blocks binding of an alternative clone, BNI8 to CTLA-4/Ig in ELISA. Mouse anti Human CD152 antibody, clone BNI3 binds to the same epitope as classified anti CTLA-4 clones 11D4 and 10A8 (Wang et al. In: Leukocyte typing VI 1997 Garland Publishing Inc. pp97-98, Bull World Health Organ. 1997). The cytoplasmic domain of CD152 contains a critical tyrosine at residue 201 phosphorylated by Janus Kinase 2 which subsequently controls surface expression through regulation of CD152 interaction with AP-2 (Shiratori et al. 1997, Chikuma et al. 2000). CD152 is expressed primarily as an intracellular antigen with transport to the cell surface under tight regulation of several molecules including Trim, PLD and TIRC7, CD152 also demonstrates rapid internalization once expressed at the cell surfac
Mouse anti Human CD158b antibody, clone GL183 recognizes human Killer cell immunoglobulin-like receptor 2DL3, also known as CD158b, KIR-023GB, MHC class I NK cell receptor, p58 natural killer cell receptor clone CL-6 or Natural killer-associated transcript 2. CD158b is a 341 amino acid, ~58 kDa single pass type-1 transmembrane glycoprotein containing two Ig-like C2-type domains. expressed by a subset of NK cells.This antibody also recognizes a ~50 kDa molecule in some NK clones, which is highly homologous to p58.2 in the extracellular domain, but has a shorter cytoplasmic tail (Moretta et al. 1985). Both molecules are members of the newly described natural killer cell receptor family.CD158b functions as a receptor specific for HLA Class I molecules, including Cw3 and related HLA-C alleles. Mouse anti Human CD158b antibody, clone GL183 can restore the lysis by human NK clones of otherwise lysis protected targets expressing Cw3.
Mouse anti Human CD154 antibody, clone TRAP-1 recognises the human CD40 ligand, also known as CD154, TNF-related activation protein (TRAP) or T-cell antigen Gp39. CD154 is a 261 amino acid ~32 kDa single pass, type-1 transmembrane glycoprotein (UniProt: P29965). CD154 is expressed on activated T lymphocytes, predominantly CD4 +ve and also on some basophils and mast cells.Mouse anti Human CD154 antibody, clone TRAP-1 binds to CD154 at an epitope distinct from the CD40 binding site (Kroczek et al. 1994).
Mouse anti Human CD104 antibody, clone 450-9D recognizes the human beta4 integrin, also known as CD104. CD104 is a ~205 kDa glycoprotein which associates with the alpha6 integrin to form the alpha6/beta4 complex. CD104 is expressed on epithelial cells, Schwann cells and various tumor cell lines. Mouse anti Human CD104 antibody, clone 450-9D recognizes an extracellular epitope on the CD104 molecule.
Mouse anti Human CD104 antibody, clone 450-9D recognizes the human beta4 integrin, also known as CD104. CD104 is a ~205 kDa glycoprotein which associates with the alpha6 integrin to form the alpha6/beta4 complex. CD104 is expressed on epithelial cells, Schwann cells and various tumor cell lines. Mouse anti Human CD104 antibody, clone 450-9D recognizes an extracellular epitope on the CD104 molecule.
Mouse anti Human CD103 antibody, clone LF61 recognizes the human CD103 cell surface antigen, a glycoprotein expressed by approximately 1% of peripheral blood lymphocytes, activated T lymphocytes and by hairy cell leukemia cells. The antigen is also expressed by intraepithelial lymphocytes. It has recently been shown to be identical to the alpha E integrin.
Mouse anti Human CD103 antibody, clone LF61 recognizes the human CD103 cell surface antigen, a glycoprotein expressed by approximately 1% of peripheral blood lymphocytes, activated T lymphocytes and by hairy cell leukemia cells. The antigen is also expressed by intraepithelial lymphocytes. It has recently been shown to be identical to the alpha E integrin.
Mouse anti Human CD103 antibody, clone LF61 recognizes the human CD103 cell surface antigen, a glycoprotein expressed by approximately 1% of peripheral blood lymphocytes, activated T lymphocytes and by hairy cell leukemia cells. The antigen is also expressed by intraepithelial lymphocytes. It has recently been shown to be identical to the alpha E integrin.
Mouse anti Human CD82 antibody, clone B-L2 recognizes human CD82, also known as C33 antigen, Metastasis suppressor Kangai-1, Suppressor of tumorigenicity 6 protein or Tetraspanin-27. CD82 is a 267 amino acid ~50-53 kDa multiple pass transmembrane glycoprotein and member of the tetraspanin family expressed by T cells, NK cells, monocytes, granulocytes and platelets.
The HIV protease (PR) hydrolyzes polyproteins of HIV virus into functional protein products that are essential for its assembly and subsequent activity. This maturation process occurs as the virion buds from the host cell. HIV protease inhibitors are used in the treatment of patients with AIDS and were considered the first breakthrough in over a decade of AIDS research. HIV protease inhibitors can lower the viral load carried by AIDS patents.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
1696
Concentration:
1 mg/ml
Storage buffer:
Phosphate buffered saline (PBS) solution with 15 mM sodium azide
Rat anti Human CD66acd antibody, clone YTH71.3 recognizes 3 members of the human CD66 family, namely CD66a, also known as Carcinoembryonic antigen-related cell adhesion molecule 1, Biliary glycoprotein 1 or CEACAM1. Clone YTH71.3 also recognizes CD66c, known as Carcinoembryonic antigen-related cell adhesion molecule 6 or Non-specific crossreacting antigen. In addition the clone also recognizes CD66d which in turn is also known as Carcinoembryonic antigen-related cell adhesion molecule 3 or CGM1 (Watt et al. 1991). CD66 family members are single pass, type 1 membrane glycoproteins bearing a single V-type Ig-like domain and a varying number of C2-type Ig-like domains. Clone YTH71.3 recognizes epitopes in the N-terminal domain of these CD66 family members.CD66 members are expressed by human neutrophils and cells of the colon where it is down regulated in some cases of colon carcinoma (Neumaier et al. 2012). CD66a is also expressed in the human alveolar adenocarcinoma cell line A549 and may play a role in Moraxella catarrhalis induced apoptosis of pulmonary epithlial cells in diseases including COPD and emphysema (N'Guessan et al. 2007).
PN-15 reacts with a lectin receptor like glycoprotein of 200 kDa (gp200), present in proximal renal tubules and on urothelium. The antigen is carbohydrate in nature. Other normal tissues that display the antigen include breast, parathyroid glands, thymus and epididymis. Among renal carcinomas 93% of primary and 84% of metastatic carcinomas are positive. Bladder cancers are also largely positive. Other tumor types include breast cancer, teratocarcinomas and parathyroid adenomas. The antigen, also called DEC-205, was assigned to CD205 at CD workshop VII. In the immune system it can facilitate tolerance to self-antigens through uptake of apoptosis derived material by dendritic cells, which in turn present fragments through MHC II and MHC I, either inducing or repressing immune responses, depending on the nature of concomitant signals.
Antibody Isotype:
IgG2b-kappa
Monosan Range:
MONOSAN
Clone:
PN-15
Concentration:
100 ug/ml
Storage buffer:
PBS with 0.02% sodium azide
Storage:
2-8°C
References 1:
Yoshida, S.O. et al, Cancer Res 49: 1802-1809 (1989)
References 2:
Li, G, et al, Anticancer Res. 20(4): 2773-8 (2000)
References 3:
Batchelder C.A. et al, Anat Rec (Hoboken) 297(8): 1392-1406 (2014)
References 4:
Cykowski M.D. et al, Ultrastruct Pathol 39(1): 69-77 (2015)
Mouse anti Human CD102 antibody, clone B-T1 recognizes human Intercellular adhesion molecule 2, also known as CD102 or ICAM-2. CD102 is a 275 amino acid ~55-65 kDa single pass type-1 transmembrane glycoprotein containing two Ig-like C2-type domains.Mouse anti Human CD102 antibody, clone B-T1 inhibits cell adhesion (Xie et al. 1995)and T cell activation and also recognizes soluble ICAM-2.
Mouse anti Human CD130 antibody, clone B-T2 recognizes soluble and membrane bound gp130 receptor, also known as CD130, Interleukin-6 receptor subunit beta, Oncostatin-M receptor subunit alpha or Interleukin-6 signal transducer. CD130 is a 918 amino acid, ~130 kDa single pass type-1 transmembrane glycoprotein containing a single Ig-like C2 type and multiple fibronectin type-III domains. CD130 can be cleaved to form a monomeric soluble ~100 kDa form of the receptor detectable in plasma and other biologic fluids where it acts as an IL-6 antagonist (Müller-Newen et al. 1998). Mouse anti Human CD130 antibody, clone B-T2 binds to an epitope dependent at least partially on the presence of the Ig-like domain (Hammacher et al. 1998).Mouse anti Human CD130 antibody, clone B-T2 has been reported to block receptor activity and inhibit IL-6 induced proliferation of XG1 cells and IL-27 mediated proliferation of naive CD4+ T cells(de Groot et al. 2012, Pflanz et al. 2004).
Mouse anti Human CD49a antibody, clone TS2/7 recognizes the human alpha 1 integrin sub-unit, which forms the VLA-1 heterodimer in association with the beta 1 integrin. VLA-1 is a receptor for collagen and laminin, and is expressed by chronically activated T cells, melanoma cells and smooth muscle cells.Mouse anti Human CD49a antibody, clone TS2/7 has been reported as being suitable for use in Western Blotting.Mouse anti Human CD49a antibody, clone TS2/7 is routinely tested in flow cytometry using HeLa cells.
Antibody Isotype:
IgG1
Monosan Range:
MONOSAN
Clone:
TS2/7
Conjugate:
Purified
Concentration:
1.0 mg/ml
Storage buffer:
PBS with 0.09% sodium azide
Storage:
2-8°C
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