H3K79me3 polyclonal antibody - Classic

Catalog Number
50 µg/42 µl
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Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 79 (H3K79me3), using a KLH-conjugated synthetic peptide.

Concentration0.76 µg/µl
Species reactivityHuman, mouse, yeast
PurityAffinity purified
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 2 μg/ChIP Fig 1, 2
ELISA 1:100 - 1:500 Fig 3
Dot Blotting 1:20,000 Fig 4
Western Blotting 1:1,000 Fig 5
Immunofluorescence 1:300 Fig 6
* 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

    ChIP-seq figure A

    ChIP-seq figure B

    ChIP-seq figure C

    ChIP-seq figure D

    Figure 1 ChIP-seq results obtained with the Diagenode antibody directed against H3K79me3
    ChIP was performed as described above using 2 μg of the Diagenode antibody against H3K79me3 (Cat. No. pAb-068-050). The IP’d DNA was analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B), in a 2 Mb region from chromosome 11 (figure 2C), and in a 100 kb region surrounding the GAPDH positive control gene (figure 2D).


    Figure 2 ChIP results obtained with the Diagenode antibody directed against H3K79me3
    ChIP was performed with the Diagenode antibody against H3K79me3 (Cat. No. pAb-068-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit and the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the GAPDH promoter (Cat. No. pp-1001-050) and for exon 2 of the inactive myoglobin gene (Cat. No. pp-1006-050). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K79me3 shows a preference for active promoters.


    Figure 3 Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against H3K79me3 (Cat. No. pAb-068-050), crude serum and flow through 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 3), the titer of the antibody was estimated to be 1:3,500.

    Dot Blot

    Figure 4 Cross reactivity tests using the Diagenode antibody directed against H3K79me3
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K79me3 (Cat. No. pAb-068-050) with peptides containing other histone modifications and the unmodified H3K79. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

    Western blot

    Figure 5 Western blot analysis using the Diagenode antibody directed against H3K79me3
    Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K79me3 (Cat. No. pAb-068-050), diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker is shown on the right; the location of the protein of interest is indicated on the left.


    Figure 5 Immunofluorescence using the Diagenode antibody directed against H3K79me3
    Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K79me3 (Cat. No. C15410068) and with DAPI. Cells were fixed with 2.5% formaldehyde for 30’ and blocked with PBS/TX-100 containing 1% BSA. Figure 6A: cells were immunofluorescently labeled with the H3K79me3 antibody (left) diluted 1:300 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right). Figure 6B and C: staining of the cells with the H3K79me3 antibody after incubation of the antibody with 2 ng/μl H4K20me3 and H3K79me3 peptide, respectively.

  • 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-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
    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
  • Documents
    Datasheet H3K79me3 C15410068 DATASHEET
    Datasheet description
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K79me3 polyclonal antibody - Classic (Diagenode Cat# C15410068 Lot# A246-0040 ). 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...

    MLL-Rearranged Acute Lymphoblastic Leukemias Activate BCL-2 through H3K79 Methylation and Are Sensitive to the BCL-2-Specific Antagonist ABT-199
    Benito JM et al.
    Targeted therapies designed to exploit specific molecular pathways in aggressive cancers are an exciting area of current research. Mixed Lineage Leukemia (MLL) mutations such as the t(4;11) translocation cause aggressive leukemias that are refractory to conventional treatment. The t(4;11) translocation produces an M...

    Degree of recruitment of DOT1L to MLL-AF9 defines level of H3K79 Di- and tri-methylation on target genes and transformation potential.
    Kuntimaddi A, Achille NJ, Thorpe J, Lokken AA, Singh R, Hemenway CS, Adli M, Zeleznik-Le NJ, Bushweller JH
    The MLL gene is a common target of chromosomal translocations found in human leukemia. MLL-fusion leukemia has a consistently poor outcome. One of the most common translocation partners is AF9 (MLLT3). MLL-AF9 recruits DOT1L, a histone 3 lysine 79 methyltransferase (H3K79me1/me2/me3), leading to aberrant gene transc...

    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...

    Promoter-exon relationship of H3 lysine 9, 27, 36 and 79 methylation on pluripotency-associated genes.
    Barrand S, Andersen IS, Collas P
    Evidence links pluripotency to a gene regulatory network organized by the transcription factors Oct4, Nanog and Sox2. Expression of these genes is controlled by epigenetic modifications on regulatory regions. However, little is known on profiles of trimethylated H3 lysine residues on coding regions of these genes in...

    Chromatin states of core pluripotency-associated genes in pluripotent, multipotent and differentiated cells.
    Barrand S, Collas P
    Oct4, Nanog and Sox2 constitute a core of transcription factors controlling pluripotency. Differentiation and reprogramming studies have unraveled a few epigenetic modifications associated in relation to the expression state of OCT4, NANOG and SOX2. There is, however, no comprehensive map of chromatin states on thes...

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