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Histone H3 di/trimethyl Lys27 antibody (mAb)
| Catalog No: 39535 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39537 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
IF |
|---|---|---|---|
| 1:500 - 1:2,000 | 1 - 3 µl per ChIP | modENCODE validated. | Not recommended. IF-validated version available: 39536. |
for Histone H3 di/trimethyl Lys27 antibody (mAb)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). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points. Histone H1 is responsible for establishing higher-order chromatin structure. 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. Lysine 27 of histone H3 can be mono-, di- or trimethylated (Histone H3 monomethyl Lys27, Histone H3 dimethyl Lys27 or H3K27me3) by different histone methyltransferases such as EZH2 or NSD3. Methylation of this residue is mainly associated with transcriptional repression.
for Histone H3 di/trimethyl Lys27 antibody (mAb)Immunogen
This antibody was raised against a peptide including trimethyl-lysine 27 of histone H3.
for Histone H3 di/trimethyl Lys27 antibody (mAb)Application Notes
This Histone H3 di/trimethyl Lys27 antibody (H3K27me2/3) has been validated for use in chromatin IP (1 - 3 µl per ChIP), ChIP-Seq & ChIP-chip (5 µl each) and Western blotting (1:500 - 1:2,000 dilution). Specificity for dimethyl and trimethyl lysine 27 was confirmed by peptide competition using peptides methylated at lysines 9 and 27 of H3. Because this modification is conserved in higher eukaryotes, a wide range of reactivity is expected.
The modENCODE and NIH Roadmap Epigenomics Mapping Consortiums have implemented rigorous standardization criteria for all assays and reagents to be used. As part of this initiative, antibody specificity testing and the ability of the antibodies to work in ChIP-Seq were assessed in a large-scale study. This Histone H3 di/trimethyl Lys27 antibody was validated for ChIP-Seq in the study (see References).
for Histone H3 di/trimethyl Lys27 antibody (mAb)Buffer
Ascites containing 30% glycerol and 0.035% sodium azide.
for Histone H3 di/trimethyl Lys27 antibody (mAb)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 di/trimethyl Lys27 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 di/trimethyl Lys27 antibody (mAb).
for Histone H3 di/trimethyl Lys27 antibody (mAb)References
This antibody has been used as described below:
Histone H3 di/trimethyl Lys27 antibody (mAb) tested by Western blot.
HeLa acid extract (10 µg per lane) was probed with Histone H3 di/trimethyl Lys27 antibody (mAb) at a 1:2,000 dilution with or without pre-incubation of antibody with 1 µM peptide containing the sequence surrounding either lysine 9 or lysine 27 of Histone H3.
Lane 1: No peptide.
Lane 2: Unmodified Lys27 peptide.
Lane 3: Monomethyl Lys27 peptide.
Lane 4: Dimethyl Lys27 peptide.
Lane 5: Trimethyl Lys27 peptide.
Lane 6: Monomethyl Lys9 peptide.
Lane 7: Dimethyl Lys9 peptide.
Lane 8: Trimethyl Lys9 peptide.
Histone H3 di/trimethyl Lys27 antibody (mAb) tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 10^6 cell equivalents per ChIP) using 10 µl of Histone H3 di/trimethyl Lys27 antibody (mAb) or 2 µg 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 the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.
Histone H3 di/trimethyl Lys27 antibody specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3 di/trimethyl Lys27 antibody was applied at a dilution of 1:500 to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 7 and the results plotted. Specificity data is shown for the most reactive peptides.
for Histone H3 di/trimethyl Lys27 antibody (mAb)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 di/trimethyl Lys27 antibody (mAb)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
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
