Histone H3K9me3 antibody (pAb)

RRID: AB_2793334
Catalog No: 39765 Format: 100 µg ¥88,000 Buy
Catalog No: 39766 Format: 10 µg ¥24,000 Buy
Catalog No: 39065 Format: 50 µg ¥52,000 Buy

Request a quote for a bulk order Request Quote


Product Review Submit a Review


Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

CUT&Tag Validated ChIP Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 10 µg per ChIP
ICC/IF: 1 µg/ml dilution
WB*: 0.5 - 2 µg/ml dilution
CUT&Tag: 1 µg per 50 µl reaction

*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

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP
CUT&Tag
WB
IF(FFPE)

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

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

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an unpurified serum version (Catalog No. 39161) of this antibody is also available.

View our Guide to Histone Modifications and Biological Function.

References

Histone H3K9me3 antibody (pAb) - 100 µg (Cat. No. 39765)

 
 
 

Histone H3K9me3 antibody (pAb) - 10 µg (Cat. No. 39766)

 
 
 

 

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 106 cell equivalents per ChIP) using 10 µg of Histone H3 trimethyl Lys9 pAb 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 immunofluorescence.
Detection of Histone H3 trimethyl Lys9 by immunofluorescence. HeLa cells were stained with Histone H3 trimethyl Lys9 antibody at a dilution of 1 µg/ml. Top panel: Histone H3 trimethyl Lys9 antibody staining. Middle panel: DAPI. Bottom panel: merge.

Histone H3 trimethyl Lys9 antibody tested by Western blot.
HeLa nuclear extract (20 µg) was probed with Histone H3 trimethyl Lys9 antibody (1 µg/ml dilution).

Histone H3 trimethyl Lys9 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of 39765 for trimethyl-lysine 9 of histone H3. Peptides corresponding to regions around major sites of histone H3 methylation were spotted onto PVDF and probed with 39765 at a dilution of 1 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row.
Top row Lane 1: unmodified Lys4. Lane 2: monomethyl Lys4. Lane 3: dimethyl Lys4. Lane 4: trimethyl Lys4. Lane 5: unmodified Lys9. Lane 6: monomethyl Lys9. Lane 7: dimethyl Lys9. Lane 8: trimethyl Lys9. Lane 9: dimethyl Lys14. Lane 10: monomethyl Lys18. Lane 11: dimethyl Lys18. Lane 12: trimethyl Lys18. Bottom row, Lane 1: Unmodified Lys23. Lane 2: Monomethyl Lys23. Lane 3: Dimethyl Lys23. Lane 4: Trimethyl Lys23. Lane 5: unmodified Lys27. Lane 6: monomethyl Lys27. Lane 7: dimethyl Lys27. Lane 8: trimethyl Lys27. Lane 9: unmodified Lys36. Lane 10: monomethyl Lys36. Lane 11: dimethyl Lys36. Lane 12: trimethyl Lys36.

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.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3K9me3 antibody (pAb).

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;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • 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;
  • NEU = Neutralizing;
  • WB = Western Blotting