Diagenode

H3K64me3 Antibody

Catalog Number
Format
Price
C15410211
50 μg
$380.00
  Bulk order



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

LotA2229P
Concentration0.93 µg/µl
Species reactivityHuman
TypePolyclonal
PurityAffinity purified
HostRabbit
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 1:100 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

    H3K64me3 Antibody ChIP Grade

    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).

    H3K64me3 Antibody ELISA validation

    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.

    H3K64me3 Antibody validated in Dot Blot

    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.

    H3K64me3 Antibody validated in 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.

  • Target Description

    Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases.

  •  Applications
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    WB
    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
    Download
  •  Safety sheets
    SDS C15410211 H3K64me3 Antibody US en Download
    SDS C15410211 H3K64me3 Antibody GB en Download
    SDS C15410211 H3K64me3 Antibody BE fr Download
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    SDS C15410211 H3K64me3 Antibody ES es Download
    SDS C15410211 H3K64me3 Antibody JP ja Download
    SDS C15410211 H3K64me3 Antibody DE de Download
  •  Publications

    How to properly cite this product in your work

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

    Using our products in your publication? Let us know!

    Dynamic association of the H3K64 trimethylation mark with genes encodingexported proteins in Plasmodium falciparum.
    Jabeena, C A et al.
    Epigenetic modifications have emerged as critical regulators of virulence genes and stage-specific gene expression in Plasmodium falciparum. However, the specific roles of histone core epigenetic modifications in regulating the stage-specific gene expression are not well understood. In this study, we report an uncon...

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