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Histone H3 trimethyl Lys36 antibody (pAb)
| Catalog No: 61101 | Format: 100 µl | $340 | Buy Now |
| Catalog No: 61102 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
|---|---|---|
| 0.1 - 1 µg/ml | 10 µg per ChIP | 4 µg per ChIP-Seq 4 µg per ChIP-chip |
for Histone H3 trimethyl Lys36 antibody (pAb)Background
Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression. The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Histone H3 is methylated at lysine 36 by the Set2 (yeast) and NSD1 (mammals) methyltransferases. Dimethylation of lysine 36 of histone H3 is involved with transcriptional elongation by RNA pol II holoenzyme and is a marker of transcribed genes.
for Histone H3 trimethyl Lys36 antibody (pAb)Immunogen
This Histone H3 trimethyl Lys36 antibody was raised against a peptide containing trimethyl Lys36 of human histone H3.
for Histone H3 trimethyl Lys36 antibody (pAb)Application Notes
This Histone H3 trimethyl Lys36 antibody (H3K36me3) has been validated for use in chromatin IP (10 µg per ChIP), ChIP-Seq & ChIP-chip (4 µg each) and Western blotting (0.1 - 1 µg/ml).
for Histone H3 trimethyl Lys36 antibody (pAb)Buffer
Purified IgG in PBS, 30% glycerol, 0.035% sodium azide.
for Histone H3 trimethyl Lys36 antibody (pAb)Positive Control
Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 trimethyl Lys36 antibody.
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
Please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3 trimethyl Lys36 antibody (pAb).
Western Blot:
Nuclear extract of HeLa cells (20 µg) probed with the Histone H3 trimethyl Lys36 antibody at a dilution of 1 µg/ml.
Chromatin IP:
ChIP performed using HeLa Chromatin (1.5 x 10^6 cell equivalents per ChIP) and 10 µg of Histone H3 trimethyl Lys36 antibody (pAb) or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for either the PABPC1 (+ 0.5) or the PABPC1 ( +18) gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.
Specificity Data:
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys36 antibody for trimethyl-lysine 36 of histone H3. Peptides corresponding to regions around major sites of histone H3 methylation were spotted onto PVDF and probed with the antibody at a dilution of 1 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row. Top panel - Lane 1: unmodified Lys4. Lane 2: monomethyl Lys4. Lane 3: dimethyl Lys4. Lane 4: trimethyl Lys4. Lane 5: unmodified Lys9. Lane 6: monomethyl Lys9. Lane 7: dimethyl Lys9. Lane 8: trimethyl Lys9. Lane 9: unmodified Lys79. Lane 10: monomethyl Lys79. Lane 11: dimethyl Lys79. Lane 12: trimethyl Lys79. Bottom panel - Lane 1: Unmodified Lys23. Lane 2: Monomethyl Lys23. Lane 3: Dimethyl Lys23. Lane 4: Trimethyl Lys23. Lane 5: unmodified Lys27. Lane 6: monomethyl Lys27. Lane 7: dimethyl Lys27. Lane 8: trimethyl Lys27. Lane 9: unmodified Lys36. Lane 10: monomethyl Lys36. Lane 11: dimethyl Lys36. Lane 12: trimethyl Lys36.
Histone H3 trimethyl Lys36 antibody (pAb) tested by ChIP-Seq.
ChIP was performed using 30 ug of chromatin from a human kidney. ChIP DNA was sequenced on the Illumina GA II and 30 million sequence tags were mapped to identify H3K36me3 binding across the genome. The image is zoomed in to show H3K36me3 binding across 5 different genes on chromosome 20.
for Histone H3 trimethyl Lys36 antibody (pAb)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Histone H3 trimethyl Lys36 antibody (pAb)Guarantee
This product is guaranteed for 6 months from date of arrival.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
