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Antibodies

Histone H3K27me2 antibody (mAb)

Clone: MABI 0324
Catalog No: 61435 Format: 100 µg $410 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG2a
Purification:None
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

for Histone H3 dimethyl Lys27 antibody (mAb) (Clone MABI 0324)Application Notes


Validated Applications:
ChIP: 2 µg per ChIP
WB: 0.5 - 2 µg/ml dilution

Published Applications:
ChIP
ChIP-Seq

for Histone H3 dimethyl Lys27 antibody (mAb) (Clone MABI 0324)Immunogen

This antibody was raised against a synthetic peptide containing dimethyl Lysine 27 of human histone H3.

for Histone H3 dimethyl Lys27 antibody (mAb) (Clone MABI 0324)Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, a PBS only version of this antibody is also available.

for Histone H3 dimethyl Lys27 antibody (mAb) (Clone MABI 0324)References

 
Histone H3K27me2 antibody (mAb) tested by ChIP.

Histone H3K27me2 antibody (mAb) tested by ChIP.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 10^6 cell equivalents per ChIP) using 2 µg of Histone H3K27me2 antibody or the equivalent amount of IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from the ChIP reaction using primer pairs specific for the Myt1-9 gene, the MyoD gene or the negative control primer pair Untr12. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3K27me2 antibody (mAb) tested by Western blot.

Histone H3K27me2 antibody (mAb) tested by Western blot.
Nuclear extract of HeLa cells (30 µg) probed with Histone H3K27me2 antibody (mAb) at a dilution of 2 µg/ml.

Histone H3K27me2 antibody (mAb) specificity tested by peptide array analysis.

Histone H3K27me2 antibody (mAb) specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3K27me2 antibody was applied at a dilution of 0.2 µg/ml 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.

for Histone H3 dimethyl Lys27 antibody (mAb) (Clone MABI 0324)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 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 (mAb) (Clone MABI 0324)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 (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H3K27me2 antibody (mAb).

for Histone H3 dimethyl Lys27 antibody (mAb) (Clone MABI 0324)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 (mAb) (Clone MABI 0324)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.

This antibody is manufactured by MAB Institute, Inc.

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 (mAb)
 

Data Thumbnails

Histone H3K27me2 antibody (mAb) tested by ChIP.

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

Histone H3K27me2 antibody (mAb) tested by Western blot.

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

Histone H3K27me2 antibody (mAb) specificity tested by peptide array analysis.

Peptide array analysis of Histone H3K27me2 antibody (mAb).
(Click image to enlarge and see details).