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Antibodies

Histone H3K27me2 antibody (pAb)

Catalog No: 39245 Format: 200 µl $410 Buy Now
Catalog No: 39246 Format: 10 µl $95 Buy Now

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Antibody Type:Polyclonal
Isotype:Serum
Purification:None
Host:Rabbit
Molecular Weight:17 kDa
Reactivity:Human, Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated

for Histone H3 dimethyl Lys27 antibody (pAb)Application Notes


Validated Applications:
ChIP-Seq: 4-6 µl per ChIP
ChIP: 5 - 10 µl per ChIP
ICC/IF: 1:200 - 1:500 dilution
WB: 1:500 - 1:1,000 dilution

Published Applications:
ChIP-Seq
ChIP
WB

for Histone H3 dimethyl Lys27 antibody (pAb)Immunogen

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

for Histone H3 dimethyl Lys27 antibody (pAb)Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39919) of this antibody that was purified by Protein A Chromatography is also available.

for Histone H3 dimethyl Lys27 antibody (pAb)References

 
Histone H3K27me2 antibody (pAb) tested by ChIP-Seq.

Histone H3K27me2 antibody (pAb) tested by ChIP-Seq.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from lung tumor chromatin and 6 µl Histone H3K27me2 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 9.1 million sequence tags were mapped to identify Histone H3K27me2 binding sites.

Histone H3K27me2 antibody (pAb) tested by ChIP.

Histone H3 dimethyl Lys27 antibody tested by ChIP.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and 50 µl of Ready-to-ChIP HeLa Chromatin (Catalog No. 53015) per ChIP. Subsequent to the ChIP reaction, DNA was purified from the immunoprecipitated chromatin and a region approximately 1000 base pairs 5′ of the transcriptional start site of the HoxD13 gene was amplified by PCR.
    Lane 1: ChIP using negative control rabbit IgG.
    Lane 2: ChIP using 10 µl of Histone H3 dimethyl Lys27 antibody.
    Lane 3: PCR input control.

Histone H3K27me2 antibody (pAb) tested by immunofluorescence.

Histone H3 dimethyl Lys27 antibody tested by immunofluorescence.
Staining of HeLa cells with Histone H3 dimethyl Lys27 antibody (1:500 dilution, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).

Histone H3K27me2 antibody (pAb) tested by Western blot.

Histone H3 dimethyl Lys27 antibody tested by Western blot.
Detection of Histone H3 dimethyl Lys27 by Western blot. The analysis was performed using Histone H3 dimethyl Lys27 antibody at a 1:1,000 dilution.
    Lane 1: HeLa acid extract (20 µg per lane).
    Lane 2: Recombinant histone H3 (200 ng).

Histone H3K27me2 antibody (pAb) tested by dot blot analysis.

Histone H3 dimethyl Lys27 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys27 antibody for dimethyl Lys27 histone H3. Methylated peptides corresponding to the immunogen and related sequences derived from histone H3 were spotted onto PVDF and probed with the antibody at a 1:5,000 dilution. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: Unmodified lysine 4 peptide. Lane 2: Monomethyl lysine 4 peptide. Lane 3: Dimethyl lysine 4 peptide. Lane 4: Trimethyl lysine 4 peptide. Lane 5: Unmodified lysine 9 peptide. Lane 6: Monomethyl lysine 9 peptide. Lane 7: Dimethyl lysine 9 peptide. Lane 8: Trimethyl lysine 9 peptide. Lane 9: Unmodified lysine 27 peptide. Lane 10: Monomethyl lysine 27 peptide. Lane 11: Dimethyl lysine 27 peptide. Lane 12: Trimethyl lysine 27 peptide.

for Histone H3 dimethyl 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 Histone H3 trimethyl Lys27) by different histone methyltransferases such as EZH2 or NSD3. Methylation of this residue is mainly associated with transcriptional repression.

for Histone H3 dimethyl Lys27 antibody (pAb)Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 dimethyl Lys27 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 H3K27me2 antibody (pAb).

for Histone H3 dimethyl 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 dimethyl 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 H3K27me2 antibody (pAb)
 

Data Thumbnails

Histone H3K27me2 antibody (pAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K27me2 pAb.
(Click image to enlarge and see details.)

Histone H3K27me2 antibody (pAb) tested by ChIP.

ChIP of Histone H3K27me2 pAb.
(Click image to enlarge and see details.)

Histone H3K27me2 antibody (pAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H3K27me2 pAb.
(Click image to enlarge and see details.)

Histone H3K27me2 antibody (pAb) tested by Western blot.

Western blot of Histone H3K27me2 pAb.
(Click image to enlarge and see details.)

Histone H3K27me2 antibody (pAb) tested by dot blot analysis.

Dot blot of Histone H3K27me2 pAb.
(Click image to enlarge and see details.)