Active Motif,
Tools to analyze nuclear function,
Your CartYour Cart 0 items
抗体

Histone H3K27me2me3 antibody (mAb)

Clone: 7B11
商品コード: 39535 Format: 100 µl ¥69,000 Buy Now
商品コード: 39537 Format: 10 µl ¥20,000 Buy Now

Request a quote for a bulk orderBulk Order Button


Antibody Type:Monoclonal
Isotype:IgG
Purification:Ascites
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Mouse, Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)Application Notes


Validated Applications:
ChIP: 1 - 3 µl per ChIP
WB: 1:500 - 1:2,000 dilution

Published Applications:
ChIP-Seq
ChIP
RNA-FISH
WB
IF
See references for more information. Individual optimization may be required.

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 di/trimethyl Lys27 antibody was validated for ChIP-Seq in the study (see reference).

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)Immunogen

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

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)Buffer

Ascites containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39536) of this antibody that was purified by Protein A Chromatography is also available.

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)References

 
Histone H3K27me2me3 antibody (mAb) tested by ChIP.

Histone H3 di/trimethyl Lys27 antibody (mAb) tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 µl of Histone H3 di/trimethyl Lys27 antibody (mAb) or 2 µg of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3K27me2me3 antibody (mAb) tested by Western blot.

Histone H3 di/trimethyl Lys27 antibody (mAb) tested by Western blot.
HeLa acid extract (10 µg per lane) was probed with Histone H3 di/trimethyl Lys27 antibody (mAb) at a 1:2,000 dilution with or without pre-incubation of antibody with 1 µM peptide containing the sequence surrounding either lysine 9 or lysine 27 of Histone H3.
    Lane 1: No peptide.
    Lane 2: Unmodified Lys27 peptide.
    Lane 3: Monomethyl Lys27 peptide.
    Lane 4: Dimethyl Lys27 peptide.
    Lane 5: Trimethyl Lys27 peptide.
    Lane 6: Monomethyl Lys9 peptide.
    Lane 7: Dimethyl Lys9 peptide.
    Lane 8: Trimethyl Lys9 peptide.

Histone H3 di/trimethyl Lys27 antibody (mAb) specificity tested by peptide array analysis.

Histone H3 di/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 di/trimethyl Lys27 antibody was applied at a dilution of 1:500 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.

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)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 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)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.

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)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 Zc3h13 (Cat. No. 71022) 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 H3K27me2me3 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 H3K27me2me3 antibody (mAb).

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 H3K27me2me3 antibody (mAb).

for Histone H3 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)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 di/trimethyl Lys27 antibody (mAb) (Clone 7B11)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 H3K27me2me3 antibody (mAb)
 

Data Thumbnails

Histone H3K27me2me3 antibody (mAb) tested by ChIP.

ChIP of Histone H3K27me2me3 mAb.
(Click image to enlarge and see details.)

Histone H3K27me2me3 antibody (mAb) tested by Western blot.

Western blot of Histone H3K27me2me3 mAb.
(Click image to enlarge and see details.)

Histone H3 di/trimethyl Lys27 antibody (mAb) specificity tested by peptide array analysis.