H4K20me3 Antibody

Catalog Number
100 µl
  Bulk order

Monoclonal antibody raised in mouse against histone H4 containing the trimethylated lysine 20 (H4K20me3), using a KLH-conjugated synthetic peptide.

Concentrationnot determined
Species reactivityHuman, zebrafish
PurityConcentrated supernatant
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 3 μl/ChIP Fig 1
Western Blotting 1:50 Fig 2
Immunofluorescence 1:200 Fig 3
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 μl per IP.
  • Validation Data

    H4K20me3 Antibody ChIP Grade

    Figure 1 ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me3
    ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me3 (cat. No. SN-148-100) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (cat. No. kch-maglow-016), using sheared chromatin from 10,000 cells. Two different quantities of antibody (3 and 12 μl per ChIP experiment) were analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter and for the inactive gene TSH2B. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H4K20me3 Antibody validated in Western Blot

    Figure 2 Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me3
    Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode monoclonal antibody against H4K20me3 (cat. No. SN-148-100) diluted 1:50 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.

    H4K20me3 Antibody validated in Immunofluorescence

    Figure 3 Immunofluorescence with the Diagenode monoclonal antibody directed against H4K20me3
    HEK293T cells were stained with the Diagenode antibody against H4K20me3 (cat. No. SN-148-100, green), with human autoantibodies against the centromeres (CREST, Cortex Biochem, pink) and with DAPI (blue). Cells were fixed with 4% paraformaldehyde and blocked with PBS containing 0.1% Tween, 2% BSA, and 5% normal goat serum. Cells were immunofluorescently labeled with the H4K20me3 antibody (diluted 1:200 in blocking solution) and the CREST antibody (diluted 1:1000 in blocking solution) by overnight incubation at 4°C. Subsequently cells were labeled with goat-anti-mouse antibody conjugated to Alexa488 and goat-anti-human antibody conjugated to Alexa633 (both diluted 1:200 in blocking solution and incubated for 1 hour at RT). Cells were mounted with Vectashield containing DAPI. Immunofluorescence wwas performed by Dr. E. Baart, Erasmus University, Rotterdam, the Netherlands.

  • 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
    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 H4K20me3 SN-148-100 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

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H4K20me3 Antibody (Diagenode Cat# C15100148 Lot# 001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Role of Annexin gene and its regulation during zebrafish caudal fin regeneration
    Saxena S, Purushothaman S, Meghah V, Bhatti B, Poruri A, Meena Lakshmi MG, Sarath Babu N, Murthy CL, Mandal KK, Kumar A, Idris MM
    The molecular mechanism of epimorphic regeneration is elusive due to its complexity and limitation in mammals. Epigenetic regulatory mechanisms play a crucial role in development and regeneration. This investigation attempted to reveal the role of epigenetic regulatory mechanisms, such as histone H3 and H4 lysine ac...

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