H3K27me3 Antibody

Catalog Number
100 µg
  Bulk order

Monoclonal antibody raised in rabbit against the region of histone H3 containing the trimethylated lysine 27 (H3K27me3), using a KLH-conjugated synthetic peptide.

Concentration1 μg/μl
Species reactivityHuman, wide range expected.
TypeMonoclonal ChIP-seq grade
PurityAffinity purified polyclonal antibody.
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 50% glycerol, 1% BSA and 0.09% azide.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 0.5 - 1 µg per IP Fig 1, 2
Dot Blotting 1:20,000 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 0.5 - 5 µg per IP.

  • Validation data
    H3K27me3 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3
    ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K27me3 Antibody ChIP-seq Grade

    H3K27me3 Antibody for ChIP-seq

    H3K27me3 Antibody for ChIP-seq assay

    H3K27me3 Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3
    ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K27me3 (cat. No. C15210017) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence of chromosome 1 and a zoomin to a 3 Mb region (fig 2A and B) and in two genomic regions surrounding the MYT1 and TSH2B positive control genes (fig 2C and D, respectively).

    H3K27me3 Antibody validated in Dot Blot

    Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3
    To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.

    H3K27me3 Antibody validated in Western blot

    Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3
    Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown 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. Trimethylation of histone H3K27 is associated with inactive genomic regions.

  •  Applications
    ChIP-qPCR (ab)
    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
  •  Documents
    Datasheet H3K27me3 monoclonal antibody DATASHEET
    Monoclonal antibody raised in rabbit against the region of histone H3 containing the trimethylate...
    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

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