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H3K79me2 Antibody - ChIP-seq Grade (sample size)

Catalog Number
Format
Price
C15410051-10
10 µg
$95.00
  Bulk order
Other format



Polyclonal antibody raised in rabbit against the region of histone H3 containing the dimethylated lysine 79 (H3K79me2), using a KLH-conjugated synthetic peptide.

LotA1193D
Concentration1.1 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/ChIP-seq * 1-2 μg/ChIP Fig 1, 2
ELISA 1:500 Fig 3
Dot Blotting 1:5,000 Fig 4
Western Blotting 1:200 Fig 5
* 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

    H3K79me2 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K79me2
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K79me2 (Cat. No. C15410051) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (cat. No. C01010020), using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 μg of antibody per ChIP experiment was analyzed. IgG (1 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K79me2 Antibody ChIP-seq Grade

    H3K79me2 Antibody for ChIP-seq

    H3K79me2 Antibody for ChIP-seq assay

    H3K79me2 Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K79me2
    ChIP was performed with 1 μg of the Diagenode antibody against H3K79me2 (Cat. No. C15410051) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 5 Mb region of chromosome 1 (figure 2A and B) and in two 300 kb regions surrounding the EIF2S3 and CCT5 positive control genes (figure 2C and D).

    H3K79me2 Antibody ELISA validation

    Figure 3. Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K79me2 (Cat. No. C15410051). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:6,600.

    H3K79me2 Antibody Dot Blot validation

    Figure 4. Cross reactivity tests using the Diagenode antibody directed against H3K79me2
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K79me2 (Cat. No. C15410051) with peptides containing other modifications and unmodified sequences of histone H3. 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 4 shows a high specificity of the antibody for the modification of interest.

    H3K79me2 Antibody validated in Western Blot

    Figure 5. Western blot analysis using the Diagenode antibody directed against H3K79me2
    Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K79me2 (Cat. No. C15410051) diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

  • 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
    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
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    IF
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  Documents
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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    Datasheet H3K79me2 C15410051 DATASHEET
    Polyclonal antibody raised in rabbit against the region of histone H3 containing the dimethylated...
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  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K79me2 Antibody - ChIP-seq Grade (sample size) (Diagenode Cat# C15410051-10 Lot# A1193D). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    DOT1L Activity Promotes Proliferation and Protects Cortical Neural Stem Cells from Activation of ATF4-DDIT3-Mediated ER Stress In Vitro
    Roidl D, Hellbach N, Bovio PP, Villarreal A, Heidrich S, Nestel S, Grüning BA, Boenisch U, Vogel T
    Growing evidence suggests that the lysine methyltransferase DOT1L/KMT4 has important roles in proliferation, survival, and differentiation of stem cells in development and in disease. We investigated the function of DOT1L in neural stem cells (NSCs) of the cerebral cortex. The pharmacological inhibition and shRNA-me...

    Anticheckpoint pathways at telomeres in yeast
    Ribeyre Cyril, Shore David
    Telomeres hide (or ‘cap’) chromosome ends from DNA-damage surveillance mechanisms that arrest the cell cycle and promote repair, but the checkpoint status of telomeres is not well understood. Here we characterize the response in Saccharomyces cerevisiae to DNA double-strand breaks (DSBs) flanked by varying amounts o...

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