Histone H3K9me3 antibody (mAb)

Clone: 2AG-6F12-H4
Catalog No: 39285 Format: 100 µl ¥70,000 Buy
Catalog No: 39286 Format: 10 µl ¥20,000 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1k
Purification:
Culture Supernatant
Host:
Mouse
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes

Validated Applications:
ICC/IF: 1:20 - 1:100 dilution
WB: 1:500 - 1:5,000 dilution*
DB: 1:5,000 dilution

*Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot.

Published Applications:
IF
WB

This antibody has not been validated in ChIP, but ChIP-validated antibodies are available for this target: 39161.

Immunogen

This Histone H3 trimethyl Lys9 antibody was raised against a peptide containing trimethyl-lysine 9 of human histone H3.

Buffer

Culture supernatant containing 30% glycerol, 0.035% sodium azide.

View our Guide to Histone Modifications and Biological Function.

References

 

Histone H3 trimethyl Lys9 mAb (Clone 2AG-6F12-H4) tested by immunofluorescence.
Staining of HeLa cells with Histone H3 trimethyl Lys9 mAb (Clone 2AG-6F12-H4) (1:250 dilution, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).

Histone H3K9me3 (Clone 2AG-6F12-H4) tested by Western blot.
HeLa cell acid extract (20 µg) probed with Histone H3K9me3 antibody at 1:1000.

Histone H3K9me3 (Clone 2AG-6F12-H4) tested by dot blot analysis.
Various methylated H3 proteins were spotted onto PVDF and probed with antibody at 1:5000 dilution. The amount of protein (picomoles) spotted is indicated next to each row.
Column 1: Histone H3 (Cat. No. 31294) .
Column 2: Histone H3K9me1 (Cat. No. 31281).
Column 3: Histone H3K9me2 (Cat. No. 31280).
Column 4: Histone H3K9me3 (Cat. No. 31586).
Column 5: Histone H3K4me3 (Cat. No. 31278).

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.

Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 trimethyl Lys9 antibody (Clone 2AG-6F12-H4).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 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
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting