Diagenode

H3K36me3 monoclonal antibody

Catalog Number
Format
Price
C15200183
(MAb-183-050)
50 μg/50 μl
$380.00
  Bulk order
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Monoclonal antibody raised in mouse against histone H3 trimethylated at lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide.

Lot001-12
Concentration1.0 µg/µl
Species reactivityHuman, rat
TypeMonoclonal
PurityProtein A purified
HostMouse
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP* 0.5 - 1 μg/ChIP Fig 1
ELISA 1:3,000 Fig 2
Dot Blotting 1:10,000
Western Blotting 1:1,000 - 1:2,000
Immunofluorescence 1:500 Fig 3

* 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

    H3K36me3 Antibody ChIP Grade

    Figure 1.ChIP results obtained with the Diagenode monoclonal antibody directed against H3K36me3
    ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K36me3 (Cat. No. C15200183) 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).

    H3K36me3 Antibody ELISA validation

    Figure 2. Cross reactivity of the Diagenode monoclonal antibody directed against H3K36me3
    To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K36me3 (Cat. No. MAb-183-050). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure 1 shows a high specificity of the antibody for the peptide containing the modification of interest.

    H3K36me3 Antibody validated in Immunofluorescence

    Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K36me3
    HeLa cells were stained with the Diagenode antibody against H3K36me3 (Cat. No. MAb-183-050) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • 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.

  •  実験手法
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    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
    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-qPCR (ab)
    Read more
  •  資料
    Datasheet H3K36me3 MAb-183-050 DATASHEET
    Datasheet description
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    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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  •  Safety sheets
    H3K36me3 antibody SDS GB en Download
    H3K36me3 antibody SDS US en Download
    H3K36me3 antibody SDS DE de Download
    H3K36me3 antibody SDS JP ja Download
    H3K36me3 antibody SDS BE nl Download
    H3K36me3 antibody SDS BE fr Download
    H3K36me3 antibody SDS FR fr Download
    H3K36me3 antibody SDS ES es Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K36me3 monoclonal antibody (Diagenode Cat# C15200183 Lot# 001-12). Click here to copy to clipboard.

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    PRDM9 Methyltransferase Activity Is Essential for Meiotic DNA Double-Strand Break Formation at Its Binding Sites.
    Diagouraga B, Clément JAJ, Duret L, Kadlec J, de Massy B, Baudat F
    The programmed formation of hundreds of DNA double-strand breaks (DSBs) is essential for proper meiosis and fertility. In mice and humans, the location of these breaks is determined by the meiosis-specific protein PRDM9, through the DNA-binding specificity of its zinc-finger domain. PRDM9 also has methyltransferase ...

    Jmjd2c/Kdm4c facilitates the assembly of essential enhancer-protein complexes at the onset of embryonic stem cell differentiation
    Tomaz R.A. et al.
    Jmjd2 H3K9 demethylases cooperate in promoting mouse embryonic stem cell (ESC) identity. However, little is known about their importance at the exit of ESC pluripotency. Here, we reveal that Jmjd2c facilitates this process by stabilising the assembly of mediator-cohesin complexes at lineage-specific enhancers. Funct...

    Reader domain specificity and lysine demethylase-4 family function
    Su Z. et al.
    The KDM4 histone demethylases are conserved epigenetic regulators linked to development, spermatogenesis and tumorigenesis. However, how the KDM4 family targets specific chromatin regions is largely unknown. Here, an extensive histone peptide microarray analysis uncovers trimethyl-lysine histone-binding preferences ...

    Inhibiting WEE1 Selectively Kills Histone H3K36me3-Deficient Cancers by dNTP Starvation
    Pfister SX et al.
    Histone H3K36 trimethylation (H3K36me3) is frequently lost in multiple cancer types, identifying it as an important therapeutic target. Here we identify a synthetic lethal interaction in which H3K36me3-deficient cancers are acutely sensitive to WEE1 inhibition. We show that RRM2, a ribonucleotide reductase subunit, ...

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