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Histone H3K27me3 antibody (pAb)

商品コード: 39155 Format: 100 µg ¥69,000 Buy Now
商品コード: 39157 Format: 10 µg ¥20,000 Buy Now

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

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

for Histone H3 trimethyl Lys27 antibody (pAb)Application Notes


Validated Applications:
ChIP: 5 - 10 µg per ChIP
ChIP-Seq: 5 µg each
ICC/IF: 2 µg/ml dilution
WB: 0.5 - 2 µg/ml dilution

Published Applications:
ChIP-Seq
ChIP
WB
IF

The modENCODE and NIH Roadmap Epigenomics Mapping Consortiums have implemented rigorous standardization criteria for all assays and reagents to be used. As part of this initiative, antibody specificity testing and the ability of the antibodies to work in ChIP-Seq were assessed in a large-scale study. This Histone H3 trimethyl Lys27 antibody was validated for ChIP-Seq in the study (see reference).

for Histone H3 trimethyl Lys27 antibody (pAb)Immunogen

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

for Histone H3 trimethyl Lys27 antibody (pAb)Buffer

Purified IgG in PBS (pH 7.5) with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an unpurified serum version (Catalog No. 39156) of this antibody is also available.

for Histone H3 trimethyl Lys27 antibody (pAb)References

 
Histone H3K27me3 antibody (pAb) tested by ChIP-Seq.

Histone H3 trimethyl Lys27 pAb tested by ChIP-Seq.
ChIP was performed using chromatin from the human iPS 19.11 cell line. ChIP DNA was sequenced on the Illumina GA II and sequence tags were mapped to identify H3K27me3 binding. The image shows H3K27me3 binding across a 140,000 bp region of the HOX gene cluster on chromosome 12.

Histone H3K27me3 antibody (pAb) tested by immunofluorescence.

Histone H3 trimethyl Lys27 antibody tested by immunofluorescence.
HeLa cells stained with Histone H3 trimethyl Lys27 antibody (1.7 µg/ml dilution).
    Top panel: Histone H3 trimethyl Lys27 antibody.
    Middle: DAPI.
    Bottom: Merge of both images.

Histone H3K27me3 antibody (pAb) tested by Western blot.

Histone H3 trimethyl Lys27 antibody tested by Western blot.
HeLa acid extract (20 µg) probed with Histone H3 trimethyl Lys27 antibody (1 µg/ml).

Histone H3 trimethyl Lys27 antibody specificity tested by peptide array analysis.

Histone H3 trimethyl Lys27 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 trimethyl Lys27 antibody was applied at a dilution of 1:5,000 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 trimethyl Lys27 peptides by the antibody blocked by phosphorylation at Ser28.
Array Data File

Histone H3K27me3 antibody (pAb) tested by dot blot analysis.

Histone H3 trimethyl Lys27 antibody 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 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row. Lane 1: Unmodified Lys4. Lane 2: Monomethyl Lys4. Lane 3: Dimethyl Lys4. Lane 4: Trimethyl Lys4. Lane 5: Monomethyl Lys9. Lane 6: Unmodified Lys9. Lane 7: Dimethyl Lys9. Lane 8: Trimethyl Lys9. Lane 9: Unmodified Lys27. Lane 10: Monomethyl Lys27. Lane 11: Dimethyl Lys27. Lane 12: Trimethyl Lys27.

for Histone H3 trimethyl Lys27 antibody (pAb)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 27 of histone H3 can be mono-, di- or trimethylated (Histone H3 monomethyl Lys27, Histone H3 dimethyl Lys27 or H3K27me3) by different histone methyltransferases such as EZH2 or NSD3. Methylation of this residue is mainly associated with transcriptional repression.

for Histone H3 trimethyl Lys27 antibody (pAb)Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 trimethyl Lys27 antibody.

for Histone H3 trimethyl Lys27 antibody (pAb)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 Positive Control Primer Set Pax-2 (Cat. No. 71020) and Mouse Negative Control Primer Set 1 (Cat. No. 71011) have 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).

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 H3K27me3 antibody (pAb).

for Histone H3 trimethyl Lys27 antibody (pAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for Histone H3 trimethyl Lys27 antibody (pAb)Guarantee

This product is guaranteed for 6 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
 

Technical Data Sheet

Histone H3K27me3 antibody (pAb)
 

Data Thumbnails

Histone H3K27me3 antibody (pAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K27me3 pAb.
(Click image to enlarge and see details.)

Histone H3K27me3 antibody (pAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H3K27me3 pAb.
(Click image to enlarge and see details.)

Histone H3K27me3 antibody (pAb) tested by Western blot.

Western blot of Histone H3K27me3 pAb.
(Click image to enlarge and see details.)

Histone H3 trimethyl Lys27 antibody specificity tested by peptide array analysis.

Peptide array analysis of Histone H3 trimethyl Lys27 antibody.
(Click image to enlarge and see details).

Histone H3K27me3 antibody (pAb) tested by dot blot analysis.

Dot blot of Histone H3K27me3 pAb.
(Click image to enlarge and see details.)