H3K64me3 polyclonal antibody - Classic

Catalog Number
50 μg/ 84 μl
  Bulk order

Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 64 (H3K64me3), using a KLH-conjugated synthetic peptide.

Concentration0.93 µg/µl
Species reactivityHuman
PurityAffinity purified
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1-2 μg per ChIP Fig 1
ELISA 1:500 Fig 2
Dot Blotting 1:5,000 Fig 3
Western Blotting 1:1,000 Fig 4
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
  • Validation Data


    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K64me3
    ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K64me3 (Cat. No. 15410211) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051) on sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the Sat2 and Sata satellite repeats, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).


    Figure 2. Determination of the antibody titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K64me3 (Cat. No. 15410211) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,500.

    Cross reactivity

    Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K64me3
    To check the specificity of the Diagenode antibody against H3K64me3 (Cat. No C15410211) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H3K64 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 3 shows a high specificity of the antibody for the modification of interest.

    Western blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K64me3
    Western blot was performed on histone extracts (30 μg) from HeLa cells using the Diagenode antibody against H3K64me3 (Cat. No. C15410211). The antibody was diluted 1:100 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right.

  • Applications
    Enzyme-linked immunosorbent assay. Read more
    Dot blotting Read more
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    ChIP-qPCR (ab)
    Read more
  • Documents
    Datasheet H3K64me3 C15410211 DATASHEET
    Datasheet description
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K64me3 polyclonal antibody - Classic (Diagenode Cat# C15410211 Lot# A2229P). Click here to copy to clipboard.

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