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Histone H3K9me3 antibody (mAb)

Clone: MABI 0319
商品コード: 61013 Format: 100 µg ¥67,000 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG2b
Purification:Protein G Chromatography
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Other (Wide Range)

Applications

ChIP Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

for Histone H3 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)Application Notes


Validated Applications:
ChIP: 2 - 10 µg per ChIP
WB: 0.5 - 2 µg/ml dilution

Published Applications:
ChIP
ICC/IF
SCAN
WB
See references for more information. Individual optimization may be required.

for Histone H3 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)Immunogen

This Histone H3 trimethyl Lys9 antibody was raised against a peptide containing trimethyl Lys9 of human Histone H3.

for Histone H3 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)Buffer

PBS pH 7.5 containing 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 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)References

 
Histone H3K9me3 antibody (mAb) tested by ChIP.

Histone H3 trimethyl Lys9 antibody tested by ChIP.
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 µg of Histone H3 trimethyl Lys9 pAb or the equivalent amount of rabbit IgG. RT-qPCR was performed on DNA purified from each of the ChIP DNA using a primer pair for the SAT2 alpha gene. Data are presented as Fold Enrichment of the antibody signal versus the negative control IgG using the ddCT method.

Histone H3K9me3 antibody (mAb) tested by Western blot.

Western blot of Histone H3 trimethyl Lys9 antibody.
HeLa nuclear extract (20 µg per lane) probed with Histone H3 trimethyl Lys9 antibody (2 µg/ml dilution).

Histone H3K9me3 antibody (mAb) tested by dot blot analysis.

Histone H3 trimethyl Lys9 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys9 antibody for trimethyl Lys9 of histone H3. Recombinant methylated histone proteins corresponding to the immunogen and related sequences were spotted onto PVDF and probed with Histone H3 trimethyl Lys9 at 2 µg/ml. The amount of protein (picomoles) spotted is indicated next to each row. Lane 1: unmodified H3 protein. Lane 2: monomethyl Lys4 protein. Lane 3: dimethyl Lys4 protein. Lane 4: trimethyl Lys4 protein. Lane 5: monomethyl Lys9 protein. Lane 6: dimethyl Lys9 protein. Lane 7: trimethyl Lys9 protein. Lane 8: monomethyl Lys27 protein. Lane 9: dimethyl Lys27 protein. Lane 10: trimethyl Lys27 protein.

for Histone H3 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)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 9 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SuvH39H1 or G9a. This methylated lysine can be demethylated by histone demethylases as JMJD1A, LSD1 or JMJD2C. Methylation of this residue is mainly associated with transcriptional repression.

for Histone H3 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 trimethyl Lys9 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 H3K9me3 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 H3K9me3 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 H3K9me3 antibody (mAb).

for Histone H3 trimethyl Lys9 antibody (mAb) (Clone MABI 0319)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 Lys9 antibody (mAb) (Clone MABI 0319)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 H3K9me3 antibody (mAb)
 

Data Thumbnails

Histone H3K9me3 antibody (mAb) tested by ChIP.

ChIP of Histone H3K9me3 mAb.
(Click image to enlarge and see details.)

Histone H3K9me3 antibody (mAb) tested by Western blot.

Western blot of Histone H3K9me3 mAb.
(Click image to enlarge and see details.)

Histone H3K9me3 antibody (mAb) tested by dot blot analysis.

Dot blot of Histone H3K9me3 mAb.
(Click image to enlarge and see details.)