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H3K9me3 polyclonal antibody - Classic

Catalog Number
100 µl
  Bulk order

Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 9 (H3K9me3), using a KLH-conjugated synthetic peptide.

Concentrationnot determined
Species reactivityHuman, mouse
PurityWhole antiserum
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP 1:5,000 Fig 1
ELISA 1:1,000 Fig 2
Dot Blotting 1:10,000 Fig 3
Western Blotting 1:750 Fig 4
Immunofluorescence 1:200 Fig 5
  • Validation Data


    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me3
    ChIP assays were performed using undifferentiated human teratocarcinoma cells (NCCIT), the Diagenode antibody against H3K9me3 (cat. No. CS-056-100) and optimized PCR primer sets for qPCR. Sheared chromatin from 10,000 cells was used per ChIP experiment. The antibody was diluted 1:5000. The pre-immune serum (PI, diluted 1:5000) was used as a negative control. Quantitative PCR was performed using primer sets for the satellite repeat Sat2 as a positive control and for the promoter of the house keeping gene c-fos, as a negative control. 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 H3K9me3 is preferably present at heterochromatin.


    Figure 2. 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 H3K9me3 (cat. No. CS-056-100). 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 1:35,000.


    Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9me3
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me3 (cat. No. CS-056-100) with peptides containing other histone modifications of histone H3 and H4. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 27 and 36 of H3, and of lysine 20 of H4. 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 1:10,000. 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 H3K9me3
    Histone (acid) extracts of NB4 (human promyelocytic leukemia) cells were analysed by Western blot using the Diagenode antibody against H3K9me3 (cat. No. CS-056-100) diluted 1:750 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest of interest is indicated on the left.


    Figure 5. Immunofluorescence analysis using the Diagenode antibody directed against H3K9me3
    NIH3T3 cells (mouse fibroblasts) were stained with the antibody against H3K9me3 (cat. No. CS-056-100) and with DAPI. Cells were formaldehyde fixed, permeabilized with TritonX100 and blocked with PBS containing 2.5% BSA.
    Figure 5A: cells were immunofluorescently labelled with the H3K9me3 antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to FITC.
    Figure 5B: staining of the nuclei with DAPI, which specifically labels DNA. Both antibody and DAPI staining are restricted to the nucleus. The dense signals obtained with both stainings characterize the distribution pattern of H3K9me3, which is linked to the transcriptionally inactive, condensed pericentric heterochromatin.

  •  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
    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-qPCR (ab)
    Read more
  •  Documents
    Datasheet H3K9me3 CS-056-100 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

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    Using our products in your publication? Let us know!

    Differential distribution of HP1 proteins after trichostatin a treatment influences chromosomal stability in HCT116 and WI-38 cells
    González-Barrios R, Soto-Reyes E, Quiroz-Baez R, Fabián-Morales E, Díaz-Chávez J, del Castillo V, Mendoza J, López-Saavedra A, Castro C, Herrera LA
    Abstract Background Heterochromatin protein 1 (HP1) is important in the establishment, propagation, and maintenance of constitutive heterochromatin, especi ally at the pericentromeric region. HP1 might participate in recruiting and directing Mis12 to the centromere during interphase, and HP1 disruption or abrogation...

    H19 lncRNA controls gene expression of the Imprinted Gene Network by recruiting MBD1.
    Monnier P, Martinet C, Pontis J, Stancheva I, Ait-Si-Ali S, Dandolo L
    The H19 gene controls the expression of several genes within the Imprinted Gene Network (IGN), involved in growth control of the embryo. However, the underlying mechanisms of this control remain elusive. Here, we identified the methyl-CpG-binding domain protein 1 MBD1 as a physical and functional partner of the H19 ...

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