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Histone H3 dimethyl Lys9 antibody (pAb)
| Catalog No: 39239 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39240 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | Immunofluorescence |
|---|---|---|
| 1:2,000 - 1:5,000 | 10 µl per ChIP | 1:250 - 1:1,000 |
for Histone H3 dimethyl 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 dimethyl Lys9 antibody (pAb)Immunogen
This histone H3 dimethyl Lys9 antibody was raised against a peptide including dimethyl-lysine 9 of histone H3.
for Histone H3 dimethyl Lys9 antibody (pAb)Application Notes
This Histone H3 dimethyl Lys9 antibody (H3K9me2) has been validated for use in Western blotting (1:2,000 - 1:5,000 dilution) and immunofluorescence (1:250 - 1:1,000 dilution) and chromatin IP (10 µl per ChIP).
for Histone H3 dimethyl Lys9 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
for Histone H3 dimethyl Lys9 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 dimethyl 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 dimethyl 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 dimethyl Lys9 antibody (pAb).
Histone H3 dimethyl Lys9 antibody tested by Western blot.
Detection of Histone H3 dimethyl Lys9 by Western blot. The analysis was performed using 5 µg HeLa acid extract and Histone H3 dimethyl Lys9 antibody at a 1:2,000 dilution.
Histone H3 dimethyl Lys9 antibody tested by immunofluorescence.
Staining of HeLa cells with Histone H3 dimethyl Lys9 antibody (1:1,000 dilution, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).
Histone H3 dimethyl Lys9 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys9 pAb for dimethyl-lysine 9 of histone H3. Decreasing amounts of 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 1:5,000.
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 dimethyl 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 10 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 dimethyl 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 dimethyl Lys9 antibody was applied at a dilution of 1:15,000 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.
for Histone H3 dimethyl 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 dimethyl 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
