Histone H3K27me2me3 antibody (mAb)

RRID: AB_2793247
Clone: 7B11
Catalog No: 39536 Format: 100 µg $445 Buy
Catalog No: 39538 Format: 10 µg $110 Buy

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

Applications

Western Blot Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ICC/IF: 0.5 - 2 µg/ml dilution
WB*: 0.5 - 2 µg/ml 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.

This antibody has been published for use in ChIP, but has not been validated by Active Motif for this application. ChIP-validated antibodies are available for this target: Catalog No. 39535.

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
IF
WB

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 di/trimethyl Lys27 antibody was raised against a peptide including trimethyl-lysine 27 of histone H3.

Buffer

Purified IgG in 10 mM sodium phosphate (pH 7.5), 150 mM NaCl, 30% glycerol, 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an ascites version (Catalog No. 39535) of this antibody is also available.

View our Guide to Histone Modifications and Biological Function.

References

 

Histone H3 di/trimethyl Lys27 antibody (mAb) tested by immunofluorescence.
Top: HeLa cells stained with Histone H3 di/trimethyl Lys27 antibody (mAb) at 1:500 dilution (red). Middle: Same cells stained with DAPI (blue). Bottom: Both images merged. Staining was carried out using MAX Stain™ Immunofluorescence Tools.

Histone H3 di/trimethyl Lys27 antibody (mAb) tested by Western blot.
HeLa acid extract (10 µg) probed with Histone H3 di/trimethyl Lys27 antibody (mAb) (1:500).

Histone H3 di/trimethyl Lys27 antibody (mAb) specificity data.
HeLa acid extract (10 µg per lane) probed with Histone H3 di/trimethyl Lys27 antibody (mAb) (1:500) with or without pre-incubation of antibody with 1 µM peptide containing the sequence surrounding either Lys9 or Lys27 of Histone H3.
Lane 1: no peptide Lane 2: unmodified Lys27 Lane 3: monomethyl-Lys27 Lane 4: dimethyl Lys27 Lane 5: trimethyl-Lys27 Lane 6: unmodified Lys9 Lane 7: monomethyl-Lys9 Lane 8: dimethyl-Lys9 Lane 9: trimethyl-Lys9.

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; it 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; they play a major role in regulating gene expression.

Histone H3K27 can be mono-, di- or trimethylated by different histone methyltransferases, such as EZH2 or NSD3. While histone methylation can be associated with transcriptional activation or repression, methylation of Lysine 27 of histone H3 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 di/trimethyl Lys27 antibody.

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;
  • WB = Western Blotting