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Histone H3 dimethyl Lys9 antibody (mAb)
| Clone: MABI 0307 |
| Catalog No: 39683 | Format: 100 µg | $340 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | Immunofluorescence |
|---|---|---|
| 0.5 - 2 µg/ml | 10 µg per ChIP | 1 - 2 µg/ml |
for Histone H3 dimethyl Lys9 antibody (mAb) (Clone MABI 0307)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 (mAb) (Clone MABI 0307)Immunogen
This Histone H3 dimethyl Lys9 antibody was raised against a synthetic peptide containing dimethyl Lys9 of human histone H3.
for Histone H3 dimethyl Lys9 antibody (mAb) (Clone MABI 0307)Application Notes
This Histone H3 dimethyl Lys9 antibody has been validated for use in Western blotting (0.5 - 2 µg / ml dilution), immunofluorescence (1 - 2 µg / ml dilution) and in chromatin IP (10 µg per ChIP). See also reference "Kimura et al, 2008").
for Histone H3 dimethyl Lys9 antibody (mAb) (Clone MABI 0307)Buffer
PBS pH 7.5, 30% glycerol, 0.035% sodium azide.
for Histone H3 dimethyl Lys9 antibody (mAb) (Clone MABI 0307)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 dimethyl Lys9 antibody (Clone MABI 0307).
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 (mAb).
Bridging Antibody to Improve ChIP, MeDIP and IP
Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H3 dimethyl Lys9 antibody (mAb).
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 (mAb).
for Histone H3 dimethyl Lys9 antibody (mAb) (Clone MABI 0307)References
This antibody was used in ChIP assays as described in Kimura, H. et al (2008) Cell Struct Func 33: 61-73.
Histone H3 dimethyl Lys9 antibody tested by Western blot.
HeLa nuclear extract (20 µg) probed with Histone H3 dimethyl Lys9 antibody (2 µg/ml dilution).
Histone H3 dimethyl Lys9 antibody tested by immunofluorescence.
Staining of HeLa cells with Histone H3 dimethyl Lys9 antibody (1 µg/ml, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).
Histone H3 dimethyl Lys9 mAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys9 mAb for dimethyl-Lys9 histone H3. Recombinant methylated proteins corresponding to the immunogen and related proteins were spotted onto PVDF and probed with the antibody at 0.3 µg/ml. The amount of protein (picomoles) spotted is indicated next to each row.
Lane 1: monomethyl lysine 9 protein.
Lane 2: dimethyl lysine 9 protein.
Lane 3: trimethyl lysine 9 protein.
No detection of proteins (unmodified, mono-, di-, or tri-methylated) corresponding to lysine 27 of histone H3 was observed with Histone H3 dimethyl Lys9 antibody.
Histone H3 dimethyl Lys9 antibody tested by ChIP.
Chromatin IP performed using the ChiP-IT Express Kit (Catalog No. 53008) and 100 µl of Ready-to-ChIP HeLa Chromatin (Catalog No. 53015) per ChIP. Subsequent to the ChIP reaction, DNA was purified from the immunoprecipitated chromatin and a region 1 kb upstream of the human HOXD13 promoter was amplified by PCR.
- Lane 1: ChIP using 10 µg of Histone H3 dimethyl Lys9 antibody.
- Lane 2: PCR input control.
- Lane 3: ChIP using negative control mouse IgG.
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 0.1 µg / ml 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 dimethyl Lys9 peptide is inhibited by dimethylation at Arg8 and blocked by Ser10 and Thr11 phosphorylation and by Lys14 acetylation.
Array Data File
for Histone H3 dimethyl Lys9 antibody (mAb) (Clone MABI 0307)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 (mAb) (Clone MABI 0307)Guarantee
This product is for research use only and is not for use in diagnostic procedures. 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
