The Biosensis AG-400-AG kit utilizes an ethidium bromide counter stain for a quick and effective way to visualize cell nuclei and cell bodies of cells while under UV illumination allowing the assessment of amyloid plaques and cell/tissue positioning as well in one step. Amylo-Glo RTD Ready to Dilute Staining reagent is designed to stain amyloid plaques in tissue sections. This novel marker has several advantages over other conventional markers such as Thioflavin S and Congo Red because of its unique chemical and spectral properties. (L. Schmued et al. (2012) J.Neuroscience Methods 209:120- 126). Using Amylo-Glo results in a very bright blue UV excitable stain under physiological conditions that will not bleed through when illuminated with other filters. Its brightness makes it ideal for low magnification quantification studies, while its unique excitation/emission profile and mild staining conditions makes it ideal for combination for multiple immunofluorescent labeling studies. Amylo-Glo RTD is compatible with fresh, frozen, and formalin-fixed immunohistochemistry or cytochemistry, and it is particularly good for confocal and multiple labeling because of its high fluorescent intensity and high resistance to photo-bleaching. Moreover because Amylo-Glo fluoresces in the UV channel, double and triple labeling experiments can be performed very easily (see protocol).
Product Type:
Staining Reagent
Format:
The reagents in the Amyloid Plaque Stain Reagent (100x) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
ICC,IHC-Frozen,IHC-Paraffin-embedded
Application Details:
Staining of amyloid plaques in human and animal tissues, see included protocol. EtBr counter stain stains nuclei and cell bodies for easy identification and spacial orientation.
Alternative Names:
AmyloGlo
Biosensis Brand:
Biosensis® RTD
Detection Method:
Fluorescence
Excitation/Emission:
Excitation Peak: 334 nm; Emission Peak: 533 nm - unbound, 438 nm when bound to amyloid. To visualize Amylo-glo in tissue, UV light is required. For example, Amylo-Glo tissue can be examined using an epifluoresent microscope with UV (Nikon UV-2A) filter cube. Excitation (325-375 nm) Emission (400-450 nm) is typical. Also note, it is not uncommon for Amylo-Glo to appear light yellow when examined by eye, yet appear a light blue color when photographed. Visualization of EtBr: Ethidium bromide has an excitation peak of 300 nm and an emission peak 595 nm. Most UV compatible filter sets can be used.
Shelf Life:
6 months after date of receipt (unopened vial).
Use:
For research use only.
Kit Components:
1 bottle containing 40 mL of 10X Amylo-Glo RTD (A-G RTD) solution 1 bottle containing 40 mL of 10X A-G RTD Ethidium Bromide (EtBr RTD) solution
Concentration:
10X
References:
L. Schmued et al. (2012) J.Neuroscience Methods 209:120, 126
Specificity:
Amyloid plaques both intraneuronal and vascular for A-G, Etbr, nuclei and cell bodies both DNA and RNA label
Purification:
Thin layer chromatography using alumina plates and a solvent system of ethanol and water (3:1) revealed the presence of two fluorescent isomers. No amount of starting material was detected.
Target:
Amyloid plaque
Research Areas:
Biosensis Stains & Tracing Reagents | Amyloid beta research
Amylo-Glo RTD Ready to Dilute Staining reagent is designed to stain amyloid plaques in tissue sections. This novel marker has several advantages over other conventional markers such as Thioflavin S and Congo Red because of its unique chemical and spectral properties. (L. Schmued et al. (2012) J.Neuroscience Methods 209:120- 126). Using Amylo-Glo results in a very bright blue UV excitable stain under physiological conditions that will not bleed through when illuminated with other filters. Its brightness makes it ideal for low magnification quantification studies, while its unique excitation/emission profile and mild staining conditions makes it ideal for combination for multiple immunofluorescent labeling studies. Amylo-Glo RTD is compatible with fresh, frozen, and formalin-fixed immunohistochemistry or cytochemistry, and it is particularly good for confocal and multiple labeling because of its high fluorescent intensity and high resistance to photo-bleaching. Moreover because Amylo-Glo fluoresces in the UV channel, double and triple labeling experiments can be performed very easily (see protocol).
Product Type:
Staining Reagent
Format:
The reagents in the Amyloid Plaque Stain Reagent (100x) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
ICC,IHC-Frozen,IHC-Paraffin-embedded
Application Details:
Staining of amyloid plaques in human and animal tissues, see included protocol
Alternative Names:
AmyloGlo
Biosensis Brand:
Biosensis® RTD
Detection Method:
Fluorescence
Excitation/Emission:
Excitation Peak: 334 nm; Emission Peak: 533 nm - unbound, 438 nm when bound to amyloid. To visualize Amylo-glo in tissue, UV light is required. For example, Amylo-Glo tissue can be examined using an epifluoresent microscope with UV (Nikon UV-2A) filter cube. Excitation (325-375 nm) Emission (400-450 nm) is typical. Also note, it is not uncommon for Amylo-Glo to appear light yellow when examined by eye, yet appear a light blue color when photographed.
Shelf Life:
6 months after date of receipt (unopened vial).
Use:
For research use only.
Kit Components:
5 mL of 100X Amylo-Glo RTD (A-G RTD) solution
Concentration:
100X
References:
L. Schmued et al. (2012) J.Neuroscience Methods 209:120, 126
Specificity:
Amyloid plaques both intraneuronal and vascular
Purification:
Thin layer chromatography using alumina plates and a solvent system of ethanol and water (3:1) revealed the presence of two fluorescent isomers. No amount of starting material was detected.
Target:
Amyloid plaque
Research Areas:
Biosensis Stains & Tracing Reagents | Amyloid beta research
Rabbit anti-Beta Amyloid Polyclonal Antibody (Unconjugated), suitable for ELISA.
Background Info:
The beta Amyloid peptide is derived from the cleavage of the Amyloid precursor protein and varies in length from 39 to 43 amino acids. Beta amyloid peptides are the major constituents of the plaques and tangles that occur in Alzheimer's disease.
Product Type:
Antibody
Antibody Type:
Polyclonal
Format:
Lyophilized
Host Animal:
Rabbit
Species Reactivity:
Human
Immunogen:
A synthetic peptide (DAEFRHDSGYEVHH) conjugated to bovine serum albumin (BSA) corresponding to amino acid sequence 1-14 of mature human beta amyloid.
Applications:
ELISA
Antibody Isotype:
Mixed
Application Details:
ELISA. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
A proprietary preparation of human amyloid beta peptide (amino acids 1-42) that was initially monomerized by HFIP-treatment and then allowed to form oligomers by the procedure described in <a href="https://www.ncbi.nlm.nih.gov/pubmed/22423893" target="_blank" class="newA">Youmans KL et al. , 2012 , followed by lyophilisation using Biosensis' proprietary stabilization procedures. The resulting oligomeric mixture has been specially designed to allow the formation of stable, oligomeric A?1-42 peptide, multimeric complexes or oligomers. The material is intended to be used as a stable and consistent standard or positive control for oligomeric ELISA assays, as well as other research applications.
Product Type:
Peptide
Format:
Lyophilized, Supplied as 2 x 500 ng vials, each containing lyophilized A? oligomers . Note that the amount of provided oligomeric protein is based on the amount of monomeric A? used to form these oligomers. The precise formation, size and number of oligomers cannot be quantified by any known method.
Applications:
ELISA
Application Details:
Use as positive control in Oligomeric A? ELISA Kit (BEK-2215): Reconstitute one vial with 1 mL of assay buffer provided in the ELISA kit. Dilute to a concentration of 0.5-1 ng/mL. At this concentration, a positive signal will be obtained within the dynamic range of the calibration curve. Use as oligomeric A-beta peptide standard in Oligomeric A? ELISA Kit (BEK-2215): Reconstitute one vial with 1 mL of assay buffer provided in the ELISA kit. Dilute to a concentration of 2 ng/mL, which represents the highest concentration of the calibration curve. Perform a 1:2 serial dilution down to 0.031 ng/mL in assay buffer. Click <a class="newA" target="_blank" href="http://www.biosensis.com/documents/enhancedinfo/PE-1750-1000_Instructions for Generating a Calibration Curve.pdf"> here for detailed instructions on generating a calibration curve with PE-1750-1000. Use as positive control in other applications: Optimal concentrations need to be determined empirically. It is recommended to reconstitute the vial with 100 - 200 µL buffer first (eg., PBS, pH 7.4), and prepare further working dilutions thereof.
Youmans KL. et al. (2012) Intraneuronal Abeta detection in 5xFAD mice by a new Abeta-specific antibody. <a class="newA" target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/22423893"> Mol Neurodegener. 2012 Mar 16;7(1):8.
Purification:
Purified
Target:
Amyloid beta, oligomeric
Accession Number:
P05067
Research Areas:
Proteins & Peptides/Amyloid Beta | Amyloid beta research
Synthetic beta-amyloid A?1-42 was monomerized by HFIP (hexafluoro-2-propanol) treatment and dried. One vial contains 50 ?g monomeric A? peptide that can be used to form solutions of unaggregated A? monomers, aggregated A? oligomers, A? fibrils and A? protein complexes according to published protocols, and used in a variety of research applications.
Product Type:
Peptide
Format:
Lyophilized.
Applications:
ELISA
Application Details:
Preparation of unaggregated A-beta1-42: Important: unaggregated A-beta has to be prepared just prior to use! 1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important : refer to the attached <a class="newA" target="_blank" href="http://www.biosensis.com/documents/enhancedinfo/PE-1749-50_Peptide Reconstitution Instructions.pdf">instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of cold Dilution Buffer to make up to 111 µL total volume and a peptide concentration of 100 µM. Vortex-spin for 3 more times 4. Final concentration of A-beta is 450 µg/mL 5. Use reconstituted peptide immediately to avoid oligomer formation Preparation of oligomeric A-beta1-42: 1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important : refer to the attached <a class="newA" target="_blank" href="http://www.biosensis.com/documents/enhancedinfo/PE-1749-50_Peptide Reconstitution Instructions.pdf">instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of cold Dilution Buffer to make up to 111 µL total volume and a peptide concentration of 100 µM 4. Vortex-spin for 3 more times 5. Incubate the solution at 2-8ºC for 24 hours (protected from light) 6. Final concentration of A-beta is 450 µg/mL 7. Once reconstituted and oligomerized, o-A-beta should be used as soon as possible and within 7 days to ensure the stability of the oligomers Note: while the concentration of monomeric A-beta peptide used to form the oligomeric complexes is accurately determined, the precise formation, size and number of oligomers cannot be quantified by any known method. Preparation of fibrillar A-beta1-42: 1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important: refer to the attached <a class="newA" target="_blank" href="http://www.biosensis.com/documents/enhancedinfo/PE-1749-50_Peptide Reconstitution Instructions.pdf">instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of 10 mM HCl to make up to 111 µL total volume and a peptide concentration of 100 µM 4. Vortex-spin for 3 more times 5. Incubate the solution at 37ºC for 24 hours (protected from light) 6. Final concentration of A-beta is 450 µg/mL Preparation of A-beta1-42 Complexes: Important: only unagreggated A-beta will form complexes. Use A-beta peptide immediately after reconstitution to form complexes. 1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important: refer to the attached <a class="newA" target="_blank" href="http://www.biosensis.com/documents/enhancedinfo/PE-1749-50_Peptide Reconstitution Instructions.pdf">instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of cold Dilution Buffer to make up to 111 µL total volume and a peptide concentration of 100 µM 4. Vortex-spin for 3 more times 5. Use reconstituted peptide immediately to avoid oligomer formation 6. Mix the A-beta monomer with its complex partner (eg., lipoprotein) at desired concentrations in PBS, pH 7.4, or other suitable buffers compatible with its intended application 7. Incubate at room temperature for 2 hours without shaking 8. Use complexes immediately after incubation These protocols are based on procedures published by <a class="newA" target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/22423893">Youmans KL et al., 2012 and <a class="newA" target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/23293020">Tai LM et al., 2013 , and we refer to these publications and other relevant literature for further details. Provided working concentrations are only meant to guide the user. Optimal concentrations depend on the experimental design and need to be determined empirically.
3 vials. 1 x 50 ?g Abeta1-42 peptide; 1 x 100 ?L Reconstituting Buffer, 1 x 1 mL Dilution Buffer
References:
Youmans KL. et al. (2012) Intraneuronal Abeta detection in 5xFAD mice by a new Abeta-specific antibody. <a class="newA" target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/22423893"> Mol Neurodegener. 2012 Mar 16;7(1):8. Tai LM. et al. (2013) Levels of Soluble Apolipoprotein E/Amyloid-beta (A?) Complex Are Reduced and Oligomeric Abeta Increased with APOE4 and Alzheimer Disease in a Transgenic Mouse Model and Human Samples. <a class="newA" target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/23293020"> J Biol Chem. 2013 Feb 22;288(8):5914-26.
Purification:
Purified
Target:
Amyloid beta, 1-42
Accession Number:
P05067
Research Areas:
Proteins & Peptides/Amyloid Beta | Amyloid beta research
The amyloid beta peptide is derived from the cleavage of the Amyloid precursor protein (APP) and varies in length from 39 to 43 amino acids. However, the form(s) of amyloid-beta peptide (A? associated with the pathology characteristic of Alzheimer's disease (AD) remains unclear. In particular, the neurotoxicity of intraneuronal A? accumulation is an area of considerable research and controversy principally because antibodies thought to be specific for A? have been shown to actually detect intraneuronal APP and not A? exclusively. MOAB-2 (mouse IgG2b) is a pan-specific, high-titer antibody to A? residues 1-4 as demonstrated by biochemical and immunohistochemical analyses (IHC), and is highly specific just to amyloid beta peptide. MOAB-2 did not detect APP or APP-CTFs in cell culture media/lysates (HEK-APPSwe or HEK APPSwe/BACE1) or in brain homogenates from transgenic mice expressing 5 familial AD (FAD) mutation (5xFAD mice). Using IHC on 5xFAD brain tissue, MOAB-2 immunoreactivity co-localized with C-terminal antibodies specific for A?40 and A?42. MOAB-2 did not co-localize with either N- or C-terminal antibodies to APP. In addition, no MOAB-2-immunreactivity was observed in the brains of 5xFAD/BACE-/- mice, although significant amounts of APP were detected by N- and C-terminal antibodies to APP, as well as by 6E10. In both 5xFAD and 3xTg mouse brain tissue, MOAB-2 co-localized with cathepsin-D, a marker for acidic organelles, further evidence for intraneuronal A?, distinct from A? associated with the cell membrane. MOAB-2 demonstrated strong intraneuronal and extra-cellular immunoreactivity in 5xFAD and 3xTg mouse brain tissues. Biosensis now offers biotinylated MOAB-2 antibody allowing more flexibility in experimental design by using the biotin-avidin/streptavidin detection method. Biotinylated MOAB-2 antibody may also help to reduce background staining in difficult-to-stain tissues and increase detection sensitivity. The ability of biotinylated MOAB-2 antibody to detect amyloid beta has been validated by IHC. Purified, non-biotinylated MOAB-2 antibody is available <a href="http://www.biosensis.com/moab-mouse-monoclonal-antibody-amyloid-beta-peptide-beta-4042-purified-p-1181.htmL">here .
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Lyophilized from PBS buffer, pH 7.4; contains no preservative.
Host Animal:
Mouse
Species Reactivity:
Human,Rat
Immunogen:
Recombinant human amyloid beta protein 42 (A?42): DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
Applications:
ELISA,ICC,IHC-Frozen,IHC-Paraffin-embedded,IP,WB
Clone number:
MOAB-2
Antibody Isotype:
IgG2b, lambda
Application Details:
The biotinylated MOAB-2 antibody has been tested by IHC (1:500 - 1:2,000 dilution) and is also expected to work in applications validated for the unlabelled antibody (M-1586-100) at same or higher dilutions: Western Blotting (WB), Immunohistochemistry (IHC), Immunohistochemistry/paraffin embedded IHC(P), Immunoprecipitation (IP), Immunofluorescence (IF), ELISA. Western Blotting: MOAB-2 has been tested in WB using purified synthetic beta-amyloid preparations and from transgenic mouse brain formic acid extracts (see Figure 1). Formic acid extraction/concentration is required for western blot detection from extracts. Suggested dilution of 1:2000-1:5,000 for WB, standard ECL detection systems. Tissue samples for the detection of beta-amyloid should be prepared as detailed in Youmans KL et al., 2011 (Journal of Neuroscience Methods 196: 51-59) for best results. Detection of beta-amyloid 40/42 in direct westerns can be difficult; Dot-blots of prepared samples are recommended as detailed in Youmans KL et al., 2012. Immunohistochemistry: Suggested dilution for biotinylated MOAB-2 in IHC is 1:500-1:2,000. Fresh frozen, 4% paraformaldehyde fixed frozen, or formalin fixed paraffin embedded tissues are all suitable. Antigen retrieval is required in fixed tissues for optimal staining. Antibody was tested on 4% paraformaldehyde/0.1% glutaraldehyde fixed frozen tissue from 3xTg and 5xFAD mice. MOAB-2 antibody detects intraneuronal and extracellular beta-amyloid in IHC and does not detect APP (Youmans KL et al., 2012). The antibody also reacts with archival formalin-fixed, paraffin-embedded tissue samples with antigen Heat Induced Epitope Retrieval (HIER). Recommended buffer for HIER is citrate, pH 6.0. Signal was weak without antigen retrieval. Immunoreactivity was observed in intraneural-amyloid deposition (plaque) in Alzheimer's brain. MOAB-2 was found to be extremely clean and with an excellent signal to noise ratio with no neuro-cellular diffusive staining. In addition, MOAB-2 demonstrated no significant differences in A-beta detection using paraffin fixed, free-floating sections (Youmans KL et al., 2012). Formic acid (FA) treatment resulted in optimal detection of both intraneuronal and extracellular A-beta compared to without FA (incubated in 88% FA 8 min, Youmans KL et al., 2012). Free floating tissue sections were permeabilized in TBS containing 0.25% Triton X-100 (TBSX; 3 x 10 min), blocked with 3% horse serum in TBSX (3 x 10 min) followed by 1% horse serum in TBSX (2 x10 min) and incubated with appropriate primary antibodies diluted in TBSX containing 1% horse serum overnight. See Youmans KL et al., 2012, for full IHC(P) protocol and method details. Immunofluorescence: For IF, suggested dilution is 1:100-1:500. The antibody was tested on 4% PFA fixed frozen tissue. Fixed tissues were washed in TBS (3 x 10 min), then incubated in 88% FA (8 min), and then permeabilized in TBSX (3 x 10 min), and blocked in TBSX containing 5% bovine serum albumin (BSA; 1 hr). Sections were subsequently incubated with appropriate primary antibodies diluted in TBSX containing 2% BSA overnight on an oscillatory rotator. Detection was via fluorescently labelled absorbed secondary antibodies (Youmans KL et al., 2012). Immunoprecipitation: For IP, the suggested dilution is 1:200 to 1:1,000 for labelled beta-amyloid using SA-coated beads as the capture vehicle, similar to the protocols employed by Youmans KL et al., 2012. ELISA: In an ELISA, a dilution of 1:50-1:1,000 is suggested. The antibody has been tested in ELISAs on synthetic beta-amyloid and tissue homogenates from beta-amyloid-Tg mice. Biosensis recommends optimal dilutions/concentrations should be determined by the end user for all applications. Dilutions provided are only meant to serve as a basic guide.
Tai LM et al (2016) "The role of APOE in cerebrovascular dysfunction."<a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/26884068">Acta Neuropathol. 2016 Feb 16. [Epub ahead of print] Tai LM et al (2013) Levels of soluble apolipoprotein E/amyloid-_ (A?) complex are reduced and oligomeric A? increased with APOE4 and Alzheimer disease in a transgenic mouse model and human samples.<a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/23293020?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum"> J Biol Chem. 2013 Feb 22;288(8):5914-26. K.L. Youmans et al (2012) Intraneuronal Abeta detection in 5xFAD mice by a new Abeta-specific antibody <a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/22423893?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum"> Mol Neurodegener. 2012 Mar 16;7(1):8. K.L. Youmans et al (2011) Amyloid-_42 alters apolipoprotein E solubility in brains of mice with five familial AD mutations <a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/21219931?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum"> J Neurosci Methods. 2011 Mar 15;196(1):51-9.
Specificity:
MOAB-2 detects preparations enriched in U-, O-, F-A?42, and U-A?40 by dot-blot, and is thus a pan-specific A? antibody. However, MOAB-2 is selective for the more neurotoxic A?42 compared to A?40. Indeed, MOAB-2 demonstrated a titration against antigen concentration, and detects A?40 at 2.5 pmol, but U-, O- and F-A?b42 at antigen concentrations as low as ~ 0.1 pmol (Youmans. KL et al., 2012; PMID: 22423893). MOAB-2 does not detect APP (Amyloid Precursor Protein). Human, rat, other species not yet tested. By Dot Blot, MOAB-2 detected rat A?40 and human A?40, albeit with less affinity than for A?42 (Youmans KL et al., 2012).
Purification:
Antibody was purified from cell culture supernatant by Protein G chromatography, biotinylated and buffer-exchanged into PBS, pH 7.4 buffer
Target:
Amyloid beta peptide (A-beta 40/42)
Accession Number:
P05067
Research Areas:
Antibodies & Conjugates/Biosensis Alzheimer's & Parkinson's | Amyloid beta research
This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques. This antibody recognises human amyloid beta and can be used to detect amyloid in ELISAs, WB and tissue sections.
Product Type:
Antibody
Antibody Type:
Recombinant
Format:
Liquid. PBS with 0.02% Proclin 300.
Host Animal:
Rabbit
Species Reactivity:
Human
Immunogen:
Amyloid-beta peptide.
Applications:
ELISA,EM,IHC,IP,WB
Clone number:
6.00E+10
Antibody Isotype:
IgG, kappa
Application Details:
This antibody recognises human amyloid beta and can be used to detect amyloid in ELISAs, WB and tissue sections.
Alternative Names:
amyloid beta-peptide, A-beta
Biosensis Brand:
Biosensis®
Conjugate:
Unconjugated
Shelf Life:
Store at 2-8?C for up to 3 months. For longer storage, aliquot and store at -20?C for up to 12 months.
Use:
For research use only.
Concentration:
1 mg/mL
Specificity:
This antibody binds to amino acid residues 1-16 of beta amyloid.
Purification:
Purified antibody, via affinity chromatography
Target:
Amyloid beta
Accession Number:
P05067
Research Areas:
Antibodies & Conjugates/Biosensis Alzheimer's & Parkinson's | Amyloid beta research
The amyloid beta peptide is derived from the cleavage of the Amyloid precursor protein (APP) and varies in length from 39 to 43 amino acids. However, the form(s) of amyloid-beta peptide (A? associated with the pathology characteristic of Alzheimer's disease (AD) remains unclear. In particular, the neurotoxicity of intraneuronal A? accumulation is an area of considerable research and controversy principally because antibodies thought to be specific for A? have been shown to actually detect intraneuronal APP and not A? exclusively. MOAB-2 (mouse IgG2b) is a pan-specific, high-titer antibody to A? residues 1-4 as demonstrated by biochemical and immunohistochemical analyses (IHC), and is highly specific just to amyloid beta peptide. MOAB-2 did not detect APP or APP-CTFs in cell culture media/lysates (HEK-APPSwe or HEK APPSwe/BACE1) or in brain homogenates from transgenic mice expressing 5 familial AD (FAD) mutation (5xFAD mice). Using IHC on 5xFAD brain tissue, MOAB-2 immunoreactivity co-localized with C-terminal antibodies specific for A?40 and A?42. MOAB-2 did not co-localize with either N- or C-terminal antibodies to APP. In addition, no MOAB-2-immunreactivity was observed in the brains of 5xFAD/BACE-/- mice, although significant amounts of APP were detected by N- and C-terminal antibodies to APP, as well as by 6E10. In both 5xFAD and 3xTg mouse brain tissue, MOAB-2 co-localized with cathepsin-D, a marker for acidic organelles, further evidence for intraneuronal A?, distinct from A? associated with the cell membrane. MOAB-2 demonstrated strong intraneuronal and extra-cellular immunoreactivity in 5xFAD and 3xTg mouse brain tissues.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Lyophilized, from a Protein A purified preparation in 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, 0.01% sodium azide, 0.1% trehalose, pH 7.2; contains 0.01% sodium azide as a preservative.
Host Animal:
Mouse
Species Reactivity:
Human,Rat
Immunogen:
Recombinant human amyloid beta protein 42 (A?42): DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
Western Blotting (WB), Immunohistochemistry (IHC), Immunohistochemistry/paraffin embedded IH(P), Immunoprecipitation (IP), Immunofluorescence (IF), ELISA. Antibody has been tested in WB using purified synthetic beta-amyloid preparations and from transgenic mouse brain formic acid extracts (see figure 1). Formic acid extraction/concentration is required for western blot detection from extracts. MOAB-2 antibody is specific for beta-amyloid and does not detect APP. Suggested dilution of 1:2000-1:5,000 for WB, standard ECL detection systems. Tissue samples for the detection of beta-amyloid should be prepared as detailed in K.L. Youmans et al. {Journal of Neuroscience Methods 196 (2011) 51-59} for best results. Detection of beta-amyloid 40/42 in direct westerns can be difficult; Dot-blots of prepared samples are recommended as detailed in Youmans. KL et al 2012. IR or fluorescent detection systems not yet tested, they but are expected to work well with higher primary antibody dilutions because of the increased sensitivity of the detection methods. Suggested dilutions for IHC are 1:50-1:1,000. Fresh frozen, 4% paraformaldehyde fixed frozen, or formalin fixed paraffin embedded tissues are all suitable. Optimal dilutions must be determined by the end user. Antigen retrieval is required in fixed tissues for optimal staining. Antibody was tested on 4% paraformaldehyde/0.1% glutaraldehyde fixed frozen tissue from 3xTg and 5xFAD mice. MOAB-2 antibody detects intraneuronal and extracellular beta-amyloid in IHC and does not detect APP {Youmans KL et al 2012}. The antibody also reacts with archival formalin-fixed, paraffin-embedded tissue samples with antigen Heat Induced Epitope Retrieval (HIER): Recommended Citrate, pH 6.0 buffer for HIER. Signal was weak without antigen retrieval. Immunoreactively was expressed in intraneural-amyloid deposition (plaque) in Alzheimer's brain. MoAB-2 was found to be extremely clean and with an excellent signal to noise ratio with no neuro-cellular diffusive staining. In addition MOAB-2 demonstrated no significant differences in A-beta detection using paraffin fixed, free-floating sections {Youmans KL et al 2012}. Formic acid (FA) treatment resulted in optimal detection of both intraneuronal and extracellular A-beta compared to without FA (incubated in 88% FA 8 min, Youmans KL et al 2012). Free floating tissue sections were permeabilized in TBS containing 0.25% Triton X-100 (TBSX; 3 x 10 min), blocked with 3% horse serum in TBSX (3 x 10 min) followed by 1% horse serum in TBSX (2 x10 min) and incubated with appropriate primary antibodies diluted in TBSX containing 1% horse serum overnight. See Youmans KL et al 2012 for full IH(P) protocol and method details. For IF, suggested dilution is 1:100-1:500. The antibody was tested on 4% PFA fixed frozen tissue. Fixed tissues were washed in TBS (3 x 10 min), then incubated in 88% FA (8 min), and then permeabilized in TBSX (3 x 10 min), and blocked in TBSX containing 5% bovine serum albumin (BSA; 1 hr). Sections were subsequently incubated with appropriate primary antibodies diluted in TBSX containing 2% BSA overnight on an oscillatory rotator. Detection was via fluorescently labelled absorbed secondary antibodies {Youmans KL et al 2012}. For IP, the suggested dilution is 1:200 to 1:1,000 for labeled beta-amyloid using Protein A/G conjugated beads as the capture vehicle {Youmans KL et al 2012}. In an ELISA, a dilution of 1:50-1:1000 is suggested. The antibody has been tested in ELISAs on synthetic beta-amyloid and tissue homogenates from beta-amyloid-Tg mice. Biosensis recommends optimal dilutions/concentrations should be determined by the end user for all applications. Dilutions provided are only meant to serve as a basic guide.
Alternative Names:
Beta-APP42; Beta-APP40; Beta-amyloid protein 42; Beta-amyloid protein 40; ABPP; APPI; Amyloid beta A4 protein;MOAB2;MOAB-2; Alzheimer's antibody;AB40;AB42;abeta
Biosensis Brand:
Biosensis®
Conjugate:
Unconjugated
Shelf Life:
12 months after date of receipt (unopened vial).
Use:
For research use only.
References:
Tai LM et al (2016) "The role of APOE in cerebrovascular dysfunction."<a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/26884068">Acta Neuropathol. 2016 Feb 16. [Epub ahead of print] K.L. Youmans et al (2012) Intraneuronal Abeta detection in 5xFAD mice by a new Abeta-specific antibody <a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/22423893?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum"> Mol Neurodegener. 2012 Mar 16;7(1):8. Tai LM et al (2013) Levels of soluble apolipoprotein E/amyloid-_ (A?) complex are reduced and oligomeric A? increased with APOE4 and Alzheimer disease in a transgenic mouse model and human samples.<a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/23293020?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum"> J Biol Chem. 2013 Feb 22;288(8):5914-26. K.L. Youmans et al (2011) Amyloid-_42 alters apolipoprotein E solubility in brains of mice with five familial AD mutations <a class="newA" target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed/21219931?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum"> J Neurosci Methods. 2011 Mar 15;196(1):51-9.
Specificity:
MOAB-2 detects preparations enriched in U-, O-, F-A?42, and U-A?40 by dot-blot, and is thus a pan-specific A? antibody. However, MOAB-2 is selective for the more neurotoxic A?42 compared to A?40. Indeed, MOAB-2 demonstrated a titration against antigen concentration, and detects A?40 at 2.5 pmol but U-, O- and FA?b42 at antigen concentrations as low as ~ 0.1 pmol {Youmans. KL et al 2012}. MOAB-2 does not detect APP (Amyloid precursor protein). Human, Rat, other species not yet tested.By Dot blot, MOAB-2 detected rat A?40 and human A?40, albeit with less affinity than for A?42. {Youmans. KL et al 2012}
Purification:
This product is a Protein A purified mouse IgG2b in 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, 0.01% sodium azide, pH 7.2.
Target:
Amyloid beta peptide (A-beta 40/42)
Accession Number:
P05067
Research Areas:
Antibodies & Conjugates/Biosensis Alzheimer's & Parkinson's | Amyloid beta research
The oligomeric form of Amyloid Beta peptide (A?, 1-42) has been closely linked to Alzheimer's Disease. Several ELISAs targeting A? have been developed; however, these ELISAs are known to cross-react with Amyloid Beta precursor protein (APP) and are poorly characterized against monomeric and oligomeric forms of the peptide. The Biosensis MOAB-2 antibody, developed by LaDu and co-workers <a href="https://pubmed.ncbi.nlm.nih.gov/22423893/" target="_blank">(Youmans K. et al., 2012) , has been shown to specifically detect A?, but not the precursor molecule APP. When utilized in ELISAs, the oligomeric form of A? peptide (o-A?) can be assayed independently of the other forms of the molecule when assayed with the MOAB-2 monoclonal antibody. The Biosensis oligomeric A? ELISA kit is a sandwich ELISA that allows the preferential quantification of oligomeric A? peptides. This kit is exclusive to Biosensis and consists of a pre-coated mouse monoclonal anti-A? capture antibody (MOAB-2), a biotinylated MOAB-2 detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a colored reaction product which is directly proportional to the concentration of o-A? present in samples and protein standards. The purpose of this kit is the in vitro qualitative measurement of oligomeric A? peptide levels in brain extracts and CSF samples from both transgenic mice and humans relative to a known o-A? standard. The inclusion of a highly validated oligomeric standard results in a unique, ready-to-use ELISA kit. This kit has been configured for research use only and is not to be used in diagnostic or clinical procedures.
Product Type:
ELISA Assay
Species Reactivity:
Human,Rat
Immunogen:
The standard in this ELISA is synthetically manufactured beta-amyloid peptide, amino acids 1-42 of human, HFIP treated and dried.The stabilized oligomeric beta amyloid 1-42 control complex is also constructed from the same synthetic peptide standard material. No animal systems were used for their manufacture.
Applications:
ELISA
Application Details:
ELISA. For the quantification of Oligomeric Amyloid-beta in CSF, Tissue Homogenates. Please download the detailed product insert for complete instructions for the successful use of this ELISA. Use only as directed.
The ELISA kit box contains 96-well pre-coated strip plate(s), protein standards, QC sample, detection reagents, wash and sample buffers, substrate buffer and detailed protocols.
References:
Youmans KL et al. http://www.ncbi.nlm.nih.gov/pubmed/21219931'> (2011) J Neurosci Methods 196(1): 51-9 Youmans KL et al. http://www.ncbi.nlm.nih.gov/pubmed/22423893'> (2012) Mol Neurodegener. 16;7:8 Tai ML et al. http://www.ncbi.nlm.nih.gov/pubmed/23293020'> (2013) J Biol Chem. 288(8): 5914-26 Good pathology summary: Viola KL, Klein WL. (2015) Amyloid _ oligomers in Alzheimer's disease pathogenesis, treatment, and diagnosis. http://www.ncbi.nlm.nih.gov/pubmed/25604547'> Acta Neuropathol. 2015 Feb;129(2):183-206
Specificity:
Human. MOAB-2 (mouse IgG2b) is a pan-specific, high-titer antibody to A? residues 1-4 and is highly specific just to amyloid beta peptide. The Biosensis o-A? Elisa detects A? oligomers as validated and described by Youmans KL et al (2012) and Rat by Combes M et al (2015). Rat.
Storage:
2-8°C
Range:
0.031 - 2 ng/mL
Sample Type:
CSF,Tissue Homogenates
Target:
Oligomeric Amyloid-beta
Accession Number:
P05067
Research Areas:
ELISA Assays & Reagents/ELISA Kits | Amyloid beta research
The oligomeric form of Amyloid Beta peptide (A?, 1-42) has been closely linked to Alzheimer's Disease. Several ELISAs targeting A? have been developed; however, these ELISAs are known to cross-react with Amyloid Beta precursor protein (APP) and are poorly characterized against monomeric and oligomeric forms of the peptide. The Biosensis MOAB-2 antibody, developed by LaDu and co-workers <a href="https://pubmed.ncbi.nlm.nih.gov/22423893/" target="_blank">(Youmans K. et al., 2012) , has been shown to specifically detect A?, but not the precursor molecule APP. When utilized in ELISAs, the oligomeric form of A? peptide (o-A?) can be assayed independently of the other forms of the molecule when assayed with the MOAB-2 monoclonal antibody. The Biosensis oligomeric A? ELISA kit is a sandwich ELISA that allows the preferential quantification of oligomeric A? peptides. This kit is exclusive to Biosensis and consists of a pre-coated mouse monoclonal anti-A? capture antibody (MOAB-2), a biotinylated MOAB-2 detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a colored reaction product which is directly proportional to the concentration of o-A? present in samples and protein standards. The purpose of this kit is the in vitro qualitative measurement of oligomeric A? peptide levels in brain extracts and CSF samples from both transgenic mice and humans relative to a known o-A? standard. The inclusion of a highly validated oligomeric standard results in a unique, ready-to-use ELISA kit. This kit has been configured for research use only and is not to be used in diagnostic or clinical procedures.
Product Type:
ELISA Assay
Species Reactivity:
Human,Rat
Immunogen:
The standard in this ELISA is synthetically manufactured beta-amyloid peptide, amino acids 1-42 of human, HFIP treated and dried.The stabilized oligomeric beta amyloid 1-42 control complex is also constructed from the same synthetic peptide standard material. No animal systems were used for their manufacture.
Applications:
ELISA
Application Details:
ELISA. For the quantification of Oligomeric Amyloid-beta in CSF, Tissue Homogenates. Please download the detailed product insert for complete instructions for the successful use of this ELISA. Use only as directed.
The ELISA kit box contains 96-well pre-coated strip plate(s), protein standards, QC sample, detection reagents, wash and sample buffers, substrate buffer and detailed protocols.
References:
Youmans KL et al. http://www.ncbi.nlm.nih.gov/pubmed/21219931'> (2011) J Neurosci Methods 196(1): 51-9 Youmans KL et al. http://www.ncbi.nlm.nih.gov/pubmed/22423893'> (2012) Mol Neurodegener. 16;7:8 Tai ML et al. http://www.ncbi.nlm.nih.gov/pubmed/23293020'> (2013) J Biol Chem. 288(8): 5914-26 Good pathology summary: Viola KL, Klein WL. (2015) Amyloid _ oligomers in Alzheimer's disease pathogenesis, treatment, and diagnosis. http://www.ncbi.nlm.nih.gov/pubmed/25604547'> Acta Neuropathol. 2015 Feb;129(2):183-206
Specificity:
Human. MOAB-2 (mouse IgG2b) is a pan-specific, high-titer antibody to A? residues 1-4 and is highly specific just to amyloid beta peptide.The Biosensis o-A? Elisa detects A? oligomers as validated and described by Youmans KL et al (2012) and Rat by Combes M et al (2015). Rat.
Storage:
2-8°C
Range:
0.031 - 2 ng/mL
Sample Type:
CSF,Tissue Homogenates
Target:
Oligomeric Amyloid-beta
Accession Number:
P05067
Research Areas:
ELISA Assays & Reagents/ELISA Kits | Amyloid beta research
beta Amyloid 1-42 Antibody detects endogenous levels of total beta Amyloid 1-42.
Blocking Peptide:
A corresponding control peptide is available for this antibody (please email tech@nktscientific.com for details)
Conjugate:
Unconjugated
Uniprot:
P05067
Format:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.
beta Amyloid 1-42 Antibody detects endogenous levels of total beta Amyloid 1-42.
Blocking Peptide:
A corresponding control peptide is available for this antibody (please email tech@nktscientific.com for details)
Conjugate:
Unconjugated
Uniprot:
P05067
Format:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.
Related products:
Amyloid beta research
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