Histone H3K79me2 antibody (pAb)

RRID: AB_2793395
Catalog No: 39923 Format: 100 µg ¥70,000 Buy
Catalog No: 39924 Format: 10 µg ¥20,000 Buy

Request a quote for a bulk order Bulk Order Button


Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated Western Blot Validated Dot Blot Validated

Application Notes


Validated Applications:
ChIP: 10 µg per ChIP
WB: 1 - 2 µg/ml dilution

The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection by Western blot. Individual optimization may be required.

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

Purified IgG in PBS (pH 7.5) with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an unpurified serum version (Catalog No. 39143) of this antibody is also available.

View our Guide to Histone Modifications and Biological Function.

References

 

Histone H3 dimethyl Lys79 antibody 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 µg of Histone H3 dimethyl Lys79 pAb or the equivalent amount 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 H3 dimethyl Lys79 antibody (pAb) tested by Western blot.
Nuclear extract of HeLa cells (20 µg) probed with Histone H3 dimethyl Lys79 antibody (1 µg/ml dilution).

Histone H3 dimethyl Lys79 antibody (pAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys79 antibody for dimethyl-Lys79 of histone H3. Peptides corresponding to the region around lysine 79 of histone H3 were spotted onto PVDF and probed with Histone H3 dimethyl Lys79 antibody at a dilution of 0.5 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row.
Top panel: Lane 1: unmodified Lys4. Lane 2: monomethyl Lys4. Lane 3: dimethyl Lys4. Lane 4: trimethyl Lys4. Lane 5: unmodified Lys9. Lane 6: monomethyl Lys9. Lane 7: dimethyl Lys9. Lane 8: trimethyl Lys9. Lane 9: unmodified Lys79. Lane 10: monomethyl Lys79. Lane 11: dimethyl Lys79. Lane 12: trimethyl Lys79.
Bottom panel: Lane 1: Unmodified Lys23. Lane 2: Monomethyl Lys23. Lane 3: Dimethyl Lys23. Lane 4: Trimethyl Lys23. Lane 5: unmodified Lys27. Lane 6: monomethyl Lys27. Lane 7: dimethyl Lys27. Lane 8: trimethyl Lys27. Lane 9: unmodified Lys36. Lane 10: monomethyl Lys36. Lane 11: dimethyl Lys36. Lane 12: trimethyl Lys36.

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 nuclear extract (Catalog No. 36010) 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