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Histone H3 trimethyl Lys9 antibody (pAb)
| Catalog No: 39161 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39162 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
IF |
|---|---|---|---|
| 1:1,000 - 1:5,000 | 2 - 5 µl per ChIP | 5 µl per ChIP-Seq 5 µl per ChIP-chip |
1:500 - 1:1,000 |
for Histone H3 trimethyl Lys9 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 9 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SuvH39H1 or G9a. This methylated lysine can be demethylated by histone demethylases as JMJD1A, LSD1 or JMJD2C. Methylation of this residue is mainly associated with transcriptional repression.
for Histone H3 trimethyl Lys9 antibody (pAb)Immunogen
This Histone H3 trimethyl Lys9 antibody was raised against a peptide including trimethyl-lysine 9 of histone H3.
for Histone H3 trimethyl Lys9 antibody (pAb)Application Notes
This Histone H3 trimethyl Lys9 antibody (H3K9me3) has been validated for use in chromatin IP (2 - 5 µl per ChIP), ChIP-Seq & ChIP-chip (5 µl each) immunofluorescence (1:500 - 1:1,000) and Western blotting (1:1,000 - 1:5,000 dilution). Specificity for trimethyl lysine 9 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. In addition, Western blot against 200 ng of recombinant histone H3 was negative.
for Histone H3 trimethyl Lys9 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
for Histone H3 trimethyl Lys9 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 trimethyl Lys9 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 Lys9 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 Lys9 antibody (pAb).
for Histone H3 trimethyl Lys9 antibody (pAb)References
This antibody has been used for chromatin immunoprecipitation (ChIP) in Rai, K. et al (2010) J Biol Chem 285: 4110-4121.
Histone H3 trimethyl Lys9 antibody tested by Western blot.
HeLa acid extract (10 µg) was probed with Histone H3 trimethyl Lys9 antibody (1:1,000 dilution).
Histone H3 trimethyl Lys9 antibody tested by ChIP analysis.
Top Panel: ChIP performed on fission yeast chromatin. Lane 1: ChIP performed using Histone H3 trimethyl Lys9 antibody (3 µl). Lane 2: Input DNA PCR control. Multiplex PCR was performed using primers specific for the fission yeast chromosome 1 centromere (CEN) and an internal PCR control using primers specific for a gene-free region (GFR).
Bottom Panel: Histone H3 trimethyl Lys9 antibody 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 3 µl of Histone H3 dimethyl Lys9 antibody 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 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 trimethyl Lys9 antibody tested by ChIP-chip.
ChIP was performed using chromatin from 3 million human colorectal adenocarcinoma cells. ChIP DNA was amplified by WGA, labeled and hybridized to a human tiling array. H3K9me3 binds broadly across the genome. The image shows two different regions. The left box shows many binding sites across a 7.8 Mb region on chromosome 10. The right box is zoomed in to show a dispersed binding pattern across the RHOT1 gene.
Histone H3 trimethyl Lys9 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 Lys9 antibody was applied at a dilution of 1:2,000 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.
Histone H3 trimethyl Lys9 antibody tested by immunofluorescence.
HeLa cells stained at 2 µg/ml with Histone H3 trimethyl Lys9 antibody. Top panel: Histone H3 trimethyl Lys9 antibody. Middle panel: DAPI. Bottom panel: merge.
for Histone H3 trimethyl Lys9 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 Lys9 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
