The 5-HIAA antibody was raised to 5-HIAA coupled to BSA with paraformaldehyde. The antibody produces moderate labeling of raphe neurons in normal rat. In rats whose serotonergic system has been activated, staining intensity is increased to a maximum label. Recommended dilutions of the antiserum are 1/200-1/400 for indirect immunofluorescence and 1/4000-1/8000 for biotin-streptavidin/HRP technique. The specificity of the antiserum was evaluated using a model system of gelatin-indole plugs by a method similar to published procedures (Schipper and Tilders, 1983). Results showed that the 5-HIAA antibody dose dependently stained 5-HIAA but did not stain any concentration of 5-HT or 5-HTP. The antiserum was also tested by pre-adsorption at 25 µg/mL with various BSA conjugates. While pre-adsorption with 5-HIAA conjugate completely eliminates immunolabeling, pre-adsorption with conjugates of 5-HT,5-HTP and dopamine had no effect on staining intensity or distribution of stain.
5-hmC | 5-hydroxymethylcytosine is a recently discovered DNA modification which results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of oxygenases. It may have an important roles distinct from that of 5-methylcytosine (5-mC). 5-hmC bases have been identified in Purkinje neurons, in granule cells and embryonic stem cells where they are present at high levels (up to 0,6% of total nucleotides in Purkinje cells.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Liquid
Storage Temp:
Store lyophilized/reconstituted at -20 °C; for long term storage -80°Cis recommened; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
No confirmed exceptions from predicted reactivity are currently known
Special application note:
Close structural similarity between 5-mC and 5-hmC makes them very difficult to expreimentally distinuish. Therefore affinity-based technology was employed to purify 5-hmC specific antibodies. This antibody is affinity purified and provided in PBS pH 7.4 with 0.05 % sodium azide.
5-hmC | 5-hydroxymethylcytosine is a recently discovered DNA modification which results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of oxygenases. It may have an important roles distinct from that of 5-methylcytosine (5-mC). 5-hmC bases have been identified in Purkinje neurons, in granule cells and embryonic stem cells where they are present at high levels (up to 0,6% of total nucleotides in Purkinje cells.
Product Type:
Antibody
Antibody Type:
Polyclonal
Format:
Liquid
Storage Temp:
Store lyophilized/reconstituted at -20 °C; for long term storage Store at -80 C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
The 5-HT Antibody was raised in rabbit against 5-HTP coupled to BSA with paraformaldehyde. The antibody has a proven maximum biotin-streptavidin/HRP staining at a 1/1000 - 1/2000 dilution in rat raphe nuclei. Optimal dilution will vary depending upon fixation, labeling technique and/or detection system; therefore, a dilution series is recommended. The specificity of the antiserum was evaluated using a model system of gelatin-indole plugs by a method similar to published procedures (Shipper and Tilders, 1983). Results showed that the 5-HTP antibody dose dependently stained 5-HTP but did not stain any concentration of 5-HT or 5-HIAA. The antiserum was also tested by pre-adsorption with indole/paraformaldehyde/BSA conjugates. Staining was completely blocked by pre-adsorption with 5-HTP conjugate and unaffected by 5-HIAA or 5-HT conjugate.
The 5-HT 1A Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 294-312 of the rat 5-HT1A receptor. The antiserum is provided as 100 µL of affinity purified serum containing 1% BSA. The antiserum demonstrates strongly positive labeling of rat cortex, arcuate and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100 - 1/200 in PBS - Bn/Av-HRP detection. Intensification methods such as nickel will approximately double the dilution factor as recommended. The antibody was characterized by immunohistochemistry and Western blot. Western blot showed a single band of approximately 45 kD. Preincubation of the antibody with an excess of the synthetic peptide blocked staining. Immunohistochemical staining of rat brain correlates well with Northern analysis, in situ hybridization and receptor autoradiography. BlastP database sequence homology searches confirmed that this sequence is unique to rat, mouse and human 5-HT1A receptors.
The 5-HT 2A Receptor Antibody was raised against a multiple antigenic peptide of an N-terminal synthetic sequence corresponding to amino acids 22-41 of rat 5HT2A receptor. The antibody is provided as 100 uL of affinity purified serum in PBS (0.02 M sodium phosphate with 0.15 M sodium chloride, pH 7.5) with 1% BSA (bovine serum albumin), and 0.02% sodium azide. The antiserum demonstrates strongly positive labeling of rat cortex, amygdala and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/300 - 1/500 in PBS/0.03% Triton X-100 - Bn/Av-HRP Technique. The addition of intensifying reagents such as nickel ammonium sulfate to the chromogen solution will approximately double the dilution factor as recommended. Immunolabeling is completely abolished by preadsorption with synthetic rat 5HT2A receptor (22-41). Immunolabeling of Western blot revealed a single band of approximately 53kD.
The ImmunoStar peptide control for 5-HT2A Receptor is intended for the immuno-adsorption of 5-HT2A Receptor antiserum, catalog number 24288. Pre-adsorption of 5-HT2A Receptor antiserum, diluted according to the antibody specification sheet, with 5 µg/ml 5-HT2A Receptor peptide immunogen following the instructions below provides complete blockage of 5-HT2A Receptor immunolabeling. The peptide is provided as 25 µg of lyophilized rat 5-HT2A Receptor, sequence 22-41. Also, this antiserum contains 0.09% sodium azide. Please read the instructions carefully before beginning the procedure.
The 5-HT 2C Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 439-460 of the rat 5-HT2C receptor coupled to KLH and bovine thyroglobulin. The ImmunoStar 5-HT2C receptor antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat choroid plexus and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are /500 - 1/1000 in PBS - Bn/Av-HRP technique. Intensification methods such as nickel will approximately double the dilution factor as recommended. The antibody was characterized by immunohistochemistry and Western blot. Western blotting revealed a single band of approximately 70 kD. Preincubation of the antibody with an excess of the synthetic peptide blocked staining. Immunohistochemical staining of rat brain correlates well with Northern analysis, in situ hybridization and receptor autoradiography. BlastP database sequence homology searches confirmed that this sequence is unique to rat, mouse and human 5-HT2C receptors.
Raised against synthetic peptide sequence corresponding to amino acids 17-34 of the rat 5-HT5A receptor coupled to carrier protein with glutaraldehyde.
Product Type:
Antibody - Antibodies
Antibody Type:
polyclonal
Format:
Liquid
Host Animal:
Rabbit
Species Reactivity:
Clam, Rat
Immunogen:
Rat 5-HT5A receptor (17-34)
Applications:
Immunohistochemistry, Immunocytochemistry, immunofluorescence, Western Blot
The 5-HT 5A Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 17-34 of the rat 5-HT5A receptor coupled to carrier protein with glutaraldehyde. The ImmunoStar 5-HT5A Receptor was quality control tested using standard immunohisto-chemical methods. The antiserum demonstrates strongly positive labeling of rat cortex and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100 - 1/300 in PBS/0.3% Triton X-100Â - Cy3 and 1/200Â - 1/400 in PBS/0.3% Triton X-100Â - Bn/Av-HRP. Intensification methods such as nickel will approximately double the dilution factor as recommended. The antibody was characterized by immunoblotting and immunohistochemistry. Immunoblots of rat brain extracts revealed the presence of two bands at molecular weights of 41 and 47 kD. The lower weight band agrees with the calculated molecular weight based on amino acid sequence. The higher weight may represent glycosylated receptor protein. Immunohistochemical staining of rat brain correlates well with Northern blot analysis and in situ hybridization studies. Immunolabeling is completely abolished by preadsorption with synthetic rat 5-HT5A receptor (17-34). BlastP database sequence homology searches indicate that the amino acid sequence is unique to rat and mouse 5-HT5A receptor.
The ImmunoStar 5HT 6-receptor antibody was quality control tested using standard immunohistochemical methods. The antiserum demonstrates significant labeling of rat cortex, amygdala and hippocampus and other areas using indirect immunofluorescent and biotin/avidin-HRP techniques. The addition of intensifying reagents such as nickel ammonium sulfate to the chromogen solution will approximately double the dilution factor as recommended. Immunolabeling is completely abolished by preadsorption with synthetic rat 5HT6 receptor (CLERPPGTPRHPPGPPLW). Immunolabeling of western blot revealed a single band of approximately 53kD.
The 5-HT7 Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 8-23 of the rat 5-HT7 receptor coupled to carrier protein with glutaraldehyde. The ImmunoStar 5-HT7 receptor antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat cortex and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100 - 1/300 in PBS - biotin/avidin-HRP Technique. Note: use of Triton X-100 or other detergents is not recommended. The antibody was characterized by immunohistochemistry. Immunohistochemical staining of rat brain correlates well with Northern blot analysis, in situ hybridization and receptor autoradiography studies. Immunolabeling is completely abolished by preadsorption with synthetic rat 5-HT7 receptor (8-23). BlastP database sequence homology searches indicate that the amino acid sequence is unique to rat 5-HT7A, 5-HT7B and 5-HT7C. There is also significant sequence overlap with the mouse and human forms.
The 5-HT BSA/Conjugate Control was prepared by cross-linking 5-HT creatinine sulfate complex to BSA with paraformaldehyde. Pre-adsorption of Serotonin antisera, diluted according to the antibody specification sheet, with 20 µg/ml Serotonin/BSA conjugate following the instructions below provides complete blockage of Serotonin immunolabeling. The conjugate is provided as 50 µg of lyophilized Serotonin creatinine sulfate coupled to BSA with paraformaldehyde.
The 5-HT Goat Rabbit Antibody was raised against serotonin coupled to BSA with paraformaldehyde. The ImmunoStar serotonin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat hypothalamus and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/400 - 1/800 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/5000 - 1/10,000 in PBS/0.3% Triton X-100 - Bn/Av-HRP Technique. Staining is completely eliminated by pretreatment of the diluted antibody with 100 µg of serotonin/BSA conjugate.
The 5-HT Rabbit Antibody was raised against serotonin coupled to BSA with paraformaldehyde. The ImmunoStar serotonin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat hypothalamus, raphe nuclei and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/1,000-1/2,000 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/20,000-1/40,000 in PBS/0.3% Triton X-100 - biotin/avidin-HRP Technique. Staining is completely eliminated by pretreatment of the diluted antibody with 25 ug of serotonin/BSA. Cross reactivity of Serotonin antisera was examined. With 5µg, 10µg and 25µg amounts the following substances did not react with Serotonin antisera diluted 1/20,000 using the Bn-SA/HRP labeling method: 5-hydroxytryptophan, 5-hydroxyindole -3- acetic acid, and dopamine.
The 5-HT Transporter was raised to a synthetic peptide corresponding to amino acids 579-599 of rat 5HT transporter coupled to KLH. The ImmunoStar serotonin (5HT) transporter was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat raphe nuclei, hypothalamus, cortex and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions for these methods are 1/800 - 1/1,000 in PBS/0.3% Triton X-100 - Cy3 and 1/10,000 - 1/15,000 in PBS/0.3% Triton X-100 - Biotin/avidin-HRP. By Western blot analysis using rat brain extracts of cortex, hypothalamus, midbrain, and hindbrain, the antibody specifically labels a single band. Immunolabeling is completely abolished by pre-adsorption with synthetic rat 5HT transporter (602-622).
The peptide control for 5-HT Transporter is intended for the immuno-adsorption of 5-HT Transporter antiserum, catalog number 24330. Pre-adsorption of 5-HT Transporter antiserum, diluted according to the antibody specification sheet, with 5 µg/ml 5-HT Transporter peptide immunogen following the instructions below provides complete blockage of 5-HT Transporter immunolabeling. The peptide is provided as 25 µg of lyophilized rat 5-HT Transporter, sequence 602-622. Please read the instructions carefully before beginning the procedure.
The 5-HT Transporter Peptide Control: rat 5-HT Transporter, sequence 579-599. Pre-adsorption of 5-HT Transporter antiserum, diluted according to the antibody specification sheet, with 5 µg/ml 5-HT Transporter peptide immunogen following the instructions below provides complete blockage of 5-HT Transporter immunolabeling. The peptide is provided as 25 µg of lyophilized rat 5-HT Transporter, sequence 602-622.
5-mC (5-methylcystosine) is a methylated form of the DNA base cytosine. Methylation may be involved in the regulation of gene transcription. Alternative names: 5 Me citidine, 5 Methycytosine, 5 Me Cytidine, methyl CpG.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Liquid
Storage Temp:
Store lyophilized/reconstituted at -20 °C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Host Animal:
Mouse
Species Reactivity:
Human
Expected Species:
Mouse, broad species range
Immunogen:
BSA-conjugated molecule: 5-methylcytosine (5-mC)
Applications:
Dot blot (Dot), Immunofluorescence (IF), immunoprecipitation (IP), FISH (FISH), MeDIP (methylated DNA Immunoprecipitation (IP)
5-mC (5-methylcystosine) is a methylated form of the DNA base cytosine. Methylation may be involved in the regulation of gene transcription. Alternative names: 5 Me citidine, 5 Methycytosine, 5 Me Cytidine, methyl CpG.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Liquid
Storage Temp:
Store at -80 C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Host Animal:
Mouse
Species Reactivity:
Human
Expected Species:
Mouse, plants, broad species range
Immunogen:
OVA-conjugated molecule: 5-methylcytosine (5-mC)
Applications:
immunofluorescence (IF), MeDIP (methylated DNA Immunoprecipitation (IP)
5-mC (5-methylcystosine) is a methylated form of the DNA base cytosine. Methylation may be involved in the regulation of gene transcription. Alternative names: 5 Me citidine, 5 Methycytosine, 5 Me Cytidine, methyl CpG.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Liquid
Storage Temp:
Store at -80 C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Host Animal:
Mouse
Species Reactivity:
Human
Expected Species:
Mouse, broad species range
Immunogen:
OVA-conjugated molecule: 5-methylcytosine (5-mC)
Applications:
Dot blot (Dot), ELISA (ELISA), Immunofluorescence (IF), MeDIP/MeDIP-seq (methylated DNA Immunoprecipitation (IP)
No confirmed exceptions from predicted reactivity are currently known
Special application note:
This antibody is purified by gel filtration and is present in PBS containing 0.05 % sodium azide. This antibody is very suitable for MeDIP/MeDIP-seq (methylated DNA immunoprecipitation) applications.
DNA methylation is a type of chemical modification of DNA that can be inherited and subsequently removed without changing the original DNA sequence. Therefore it is part of the epigenetic code and is also the most well characterized epigenetic mechanism. DNA methylation results in addition of a methyl group to DNA — for example, to the number 5 carbon of the cytosine pyrimidine ring — which involves reduction in gene expression. In adult somatic tissues, DNA methylation typically occurs in a CpG dinucleotide context; non-CpG methylation is prevalent in embryonic stem cells.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Liquid
Storage Temp:
Store at -20 °C; make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Purified IgM in PBS. Contains 50 % glycerol, filter sterilized.
Selected references:
Sharif et al. (2007) The SRA protein Np95 mediates epigenetic inheritance by recruiting Dnmt1 to methylated DNA. Nature. 2007 Dec 6;450(7171):908-12. doi: 10.1038/nature06397. Epub 2007 Nov 11. PMID: 17994007.Nishiyama et al. (2002) A chloroplast-resident DNA methyltransferase is responsible for hypermethylation of chloroplast genes in Chlamydomonas maternal gametes. Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):5925-30. doi: 10.1073/pnas.082120199. PMID: 11983892; PMCID: PMC122878.Sano, Imokawa & Sager (1988) Detection of heavy methylation in human repetitive DNA subsets by a monoclonal antibody against 5-methylcytosine. Biochim Biophys Acta. 1988 Nov 10;951(1):157-65. doi: 10.1016/0167-4781(88)90036-x. PMID: 2847796.Sano, Royer & Sager (1980) Identification of 5-methylcytosine in DNA fragments immobilized on nitrocellulose paper. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3581-5. doi: 10.1073/pnas.77.6.3581. PMID: 6251470; PMCID: PMC349661.
DNA methylation is a type of chemical modification of DNA that can be inherited and subsequently removed without changing the original DNA sequence. Therefore it is part of the epigenetic code and is also the most well characterized epigenetic mechanism. DNA methylation results in addition of a methyl group to DNA — for example, to the number 5 carbon of the cytosine pyrimidine ring — which involves reduction in gene expression. In adult somatic tissues, DNA methylation typically occurs in a CpG dinucleotide context; non-CpG methylation is prevalent in embryonic stem cells.
Product Type:
Antibody
Antibody Type:
Monoclonal
Format:
Liquid
Storage Temp:
Store at -20 °C; make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
5-methylcytosine (5-mC) is formed from the DNA methylation of the 5-carbon found on the cytosine ring. A 5-methylcytosine monoclonal antibody is a useful tool in identifying and discriminating between the unmodified cytosine base (C) and the methylated cytosine base (5-mC) as part of DNA methylation studies. DNA methylation plays an important role in the repression of transcription in the genome. When present in promoter regions, 5-mC is associated with stable transcriptional silencing which results in inactivation of gene function, thereby having an important role in tumorigenesis.
5-methylcytosine (5-mC) is formed from the DNA methylation of the 5-carbon found on the cytosine ring. A 5-methylcytosine monoclonal antibody is a useful tool in identifying and discriminating between the unmodified cytosine base (C) and the methylated cytosine base (5-mC) as part of DNA methylation studies. DNA methylation plays an important role in the repression of transcription in the genome. When present in promoter regions, 5-mC is associated with stable transcriptional silencing which results in inactivation of gene function, thereby having an important role in tumorigenesis.
Tris Buffer pH7.6 with BSA, and sodium azide as preservative
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