Histone H3K79me2 antibody (pAb)

RRID: AB_2561018
Catalog No: 39143 Format: 100 µl ¥70,000 Buy
Catalog No: 39144 Format: 10 µl ¥20,000 Buy

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

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Dot Blot Validated

Application Notes


Validated Applications:
ChIP: 10 µl per ChIP
ChIP-Seq: 5 µl each
WB: 1:500 - 1:5,000 dilution

Published Applications:
ChIP-Seq
ChIP
WB
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 dimethyl Lys79 antibody was validated for ChIP-Seq in the study (see reference).

Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

Immunogen

This Histone H3 dimethyl Lys79 antibody was raised against a peptide including dimethyl-lysine 79 of histone H3.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39923) of this antibody that was purified by Protein A Chromatography is also available.

View our Guide to Histone Modifications and Biological Function.

References

 

Histone H3K79me2 antibody (pAb) tested by ChIP-chip.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 3 million HeLa cells and 5 µl of antibody. ChIP DNA was amplified by WGA, labeled and hybridized to a human tiling array. The images are characteristic of the binding patterns seen at many genes across the genome with the highest binding occurring at the promoter and a gradual decrease as signal moves into the gene body.

Histone H3 dimethyl Lys79 antibody (pAb) 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 dimethyl Lys79 antibody or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on ChIP DNA using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal compared to the negative control IgG (which has been normalized to 1.0) using the ddCT method.

Histone H3 dimethyl Lys79 antibody tested by Western blot.
HeLa acid extract (10 µg) was probed with Histone H3 dimethyl Lys79 antibody (1:2,500 dilution).

Histone H3 dimethyl Lys79 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys79 antibody for dimethyl lysine 79 of histone H3. Decreasing amounts of peptides corresponding to the region around lysine 79 of histone H3 were spotted onto PVDF and probed with the antibody at a 1:10,000 dilution.
    Lane 1: Unmodified Lys79 peptide.
    Lane 2: Monomethyl Lys79 peptide.
    Lane 3: Dimethyl Lys79 peptide.
    Lane 4: Trimethyl Lys79 peptide.

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). 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.

Lysine 79 of histone H3 can be mono-, di- or trimethylated by Dot1 methylase; methylation at this residue acts as a marker of inactive chromatin regions that is critical for transcriptional silencing, and it is thought that silencing proteins such as Sir3 function by blocking Dot1 methylation.

Positive Control

Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 dimethyl Lys79 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 H3K79me2 antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 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