Histone H3K27me3 antibody (pAb)

RRID: AB_2636821
Catalog No: 39156 Format: 100 µl $455 Buy
Catalog No: 39158 Format: 10 µl $110 Buy

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Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Human, Mouse, Wide Range Predicted

Applications

CUT&RUN Validated ChIP Validated Western Blot Validated Immunoprecipitation Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 10 µl per ChIP
ICC/IF: 1:1,000 - 1:2,500 dilution
WB*: 1:1,000 - 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.

For H3K27me3, we also offer AbFlex® H3K27me3 Recombinant Antibody (rAb). For details, see Catalog No. 91167.

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:

CUT&RUN
ChIP-qPCR, ChIP-Seq
IP
WB
ICC/IF
ELISA

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

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39155) of this antibody that was purified by Protein A Chromatography is also available.

View our Guide to Histone Modifications and Biological Function.

 

Histone H3K27me3 antibody (pAb) tested by ChIP.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 15 µg of chromatin from HeLa cells and 5 µl H3K27me3 antibody. ChIP DNA was used in qPCR with the control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

Histone H3K27me3 antibody (pAb) tested by immunofluorescence.
Staining of HeLa cells with Histone H3 trimethyl Lys27 antibody (1:1,000, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).

Histone H3K27me3 antibody (pAb) tested by Western blot.
HeLa acid extract (10 µg) was probed with Histone H3 trimethyl Lys27 antibody (1:5,000 dilution).

Histone H3K27me3 antibody (pAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys27 antibody for trimethyl-lysine 27 of histone H3. 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. The amount of peptide (in picomoles) spotted is indicated next to each row.
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.

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 trimethyl Lys27 antibody.

qPCR Primers

Active Motif's Human Positive Control Primer Set MYT1 (Cat. No. 71007) and Human Negative Control Primer Set 3 (Cat. No. 71023) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Negative Control Primer Set 1 (Cat. No. 71011) has been validated for both qPCR and endpoint PCR.

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 H3K27me3 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