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Histone H3 trimethyl Lys27 antibody (pAb)
| Catalog No: 39155 | Format: 200 µg | $340 | Buy Now |
| Catalog No: 39157 | Format: 10 µg | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
IF |
|---|---|---|---|
| 0.5 - 2 µg/ml | 5 - 10 µl per ChIP | modENCODE validated. | 1.7 µg/ml |
for Histone H3 trimethyl Lys27 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). 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 trimethyl Lys27 antibody (pAb)Immunogen
This Histone H3 trimethyl Lys27 antibody was raised against a peptide including trimethyl-lysine 27 of histone H3.
for Histone H3 trimethyl Lys27 antibody (pAb)Application Notes
This Histone H3 trimethyl Lys27 antibody (H3K27me3) has been validated for use in chromatin IP (5 - 10 μl per ChIP), ChIP-Seq & ChIP-chip (5 µg each), Western blotting (0.5 - 2 μg/ml) and immunofluorescence (1.7 μg/ml). Specificity for trimethyl lysine 27 was confirmed by dot blot with non-modified histone H3 peptide or peptides mono-, di- or trimethylated at lysine 4; peptides mono-, di- or trimethylated at lysine 9; peptides mono-, di- or trimethylated at lysine 27.
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 trimethyl Lys27 antibody was validated for ChIP-Seq in the study (see References).
for Histone H3 trimethyl Lys27 antibody (pAb)Buffer
Purified rabbit IgG in PBS, pH 7.5, 30% glycerol, 0.035% sodium azide.
for Histone H3 trimethyl Lys27 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 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 trimethyl Lys27 antibody (pAb).
Compatible Secondary Antibodies
Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Histone H3 trimethyl Lys27 antibody (pAb).
for Histone H3 trimethyl Lys27 antibody (pAb)References
This antibody has been used as described below:
Histone H3 trimethyl Lys27 antibody tested by Western blot.
HeLa acid extract (20 µg) probed with Histone H3 trimethyl Lys27 antibody (1 µg/ml).
Histone H3 trimethyl Lys27 antibody tested by immunofluorescence.
HeLa cells stained with Histone H3 trimethyl Lys27 antibody (1.7 µg/ml dilution).
Top panel: Histone H3 trimethyl Lys27 antibody.
Middle: DAPI.
Bottom: Merge of both images.
Histone H3 trimethyl Lys27 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys27 antibody for trimethyl-lysine 27 of histone H3. Peptides corresponding to regions around major sites of histone H3 methylation (lysine 4, lysine 9, lysine 27) were spotted onto PVDF and probed with the antibody at a dilution of 0.5 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row.
Lane 1: Unmodified Lys4 peptide.
Lane 2: Monomethyl Lys4 peptide.
Lane 3: Dimethyl Lys4 peptide.
Lane 4: Trimethyl Lys4 peptide.
Lane 5: Monomethyl Lys9 peptide.
Lane 6: Unmodified Lys9 peptide.
Lane 7: Dimethyl Lys9 peptide.
Lane 8: Trimethyl Lys9 peptide.
Lane 9: Unmodified Lys27 peptide.
Lane 10: Monomethyl Lys27 peptide.
Lane 11: Dimethyl Lys27 peptide.
Lane 12: Trimethyl Lys27 peptide.
Histone H3 trimethyl Lys27 pAb tested by ChIP-Seq.
ChIP was performed using chromatin from the human iPS 19.11 cell line. ChIP DNA was sequenced on the Illumina GA II and sequence tags were mapped to identify H3K27me3 binding. The image shows H3K27me3 binding across a 140,000 bp region of the HOX gene cluster on chromosome 12.
Histone H3 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 trimethyl Lys27 antibody was applied at a dilution of 1:5,000 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 and those related to the immunogen. Recognition of the H3 trimethyl Lys27 peptides by the antibody blocked by phosphorylation at Ser28.
Array Data File
for Histone H3 trimethyl Lys27 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 Lys27 antibody (pAb)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
