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H3K36me2 Antibody - ChIP-seq Grade

Catalog Number
Format
Price
C15310127
(CS-127-100)
100 µl
$325.00
  Bulk order
Other format



Polyclonal antibody raised in rabbit against histone H3 containing the dimethylated lysine 36 (H3K36me2), using a KLH-conjugated synthetic peptide.

LotA239-001
ConcentrationNot determined
Species reactivityHuman, mouse, yeast: positive. Other species: not tested.
TypePolyclonal
PurityWhole antiserum from rabbit.
HostRabbit
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05% 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 µl/ChIP Fig 1, 2
ELISA 1:1,000 Fig 3
Dot Blotting 1:100,000 Fig 4
Western Blotting 1:1,000 Fig 5
Immunofluorescence 1:500 Fig 6


*
Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-10 µl per IP.

  • Validation Data

    H3K36me2 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K36me2
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K36me2 (Cat. No. C15310127) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. C01010070), using sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 5, and 10 µl per ChIP experiment was analysed. Additionally, the same titration was analysed after incubation of the antibody with 5 nmol blocking peptide (Cat. No. C16000127 ) for 1 hour at room temperature. IgG (5 µg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes GAPDH and ALDOA and for the coding region of the myogenic differentiation gene (MYOD). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K36me2 Antibody ChIP-seq Grade

    H3K36me2 Antibody for ChIP-seq

    H3K36me2 Antibody for ChIP-seq assay

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me2
    ChIP was performed with 0.5 µl of the Diagenode antibody against H3K36me2 (Cat. No. C15310127) on sheared chromatin from 1 million HeLa cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The IP’d DNA was subsequently 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 signal distribution along 3 genomic regions of chromosome 20, 12 and X, respectively.

    H3K36me2 Antibody ELISA validation

    Figure 3. Determination of the titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me2 (Cat. No. C15310127). 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 1:31,000.

    H3K36me2 Antibody validated in Dot Blot

    Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me2
    A dot blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me2 (Cat. No. C15310127) with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

    H3K36me2 Antibody validated in Western Blot

    Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me2
    Histone extracts of HeLa cells (15 µg) were analysed by Western blot using the Diagenode antibody against H3K36me2 (Cat. No. C15310127) diluted 1:1,000 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. The result of the Western analysis with the antibody is shown in lane 1; lane 2 shows the same analysis after incubation of the antibody with 5 nmol blocking peptide (Cat. No. C16000127 ) for 1 hour at room temperature.

    H3K36me2 Antibody validated in Immunofluorescence

    Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K36me2
    HeLa cells were stained with the Diagenode antibody against H3K36me2 (Cat. No. C15310127) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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.

  •  Applications
    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-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  Documents
    Datasheet H3K36me2 CS-127-100 DATASHEET
    Datasheet description
    Download
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
  •  Safety sheets
    H3K36me2 SDS GB en Download
    H3K36me2 SDS US en Download
    H3K36me2 SDS DE de Download
    H3K36me2 SDS JP ja Download
    H3K36me2 SDS BE nl Download
    H3K36me2 polyclonal antibody SDS BE fr Download
    H3K36me2 SDS FR fr Download
    H3K36me2 SDS ES es Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K36me2 Antibody - ChIP-seq Grade (Diagenode Cat# C15310127 Lot# A239-001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Histone H3 lysine 36 methyltransferase mobilizes NER factors to regulate tolerance against alkylation damage in fission yeast.
    Lim KK, Nguyen TTT, Li AY, Yeo YP, Chen ES
    The Set2 methyltransferase and its target, histone H3 lysine 36 (H3K36), affect chromatin architecture during the transcription and repair of DNA double-stranded breaks. Set2 also confers resistance against the alkylating agent, methyl methanesulfonate (MMS), through an unknown mechanism. Here, we show that Schizosa...

    Miz1 Controls Schwann Cell Proliferation via H3K36me2 Demethylase Kdm8 to Prevent Peripheral Nerve Demyelination
    Fuhrmann D. et al.
    Schwann cell differentiation and myelination depends on chromatin remodeling, histone acetylation, and methylation, which all affect Schwann cell proliferation. We previously reported that the deletion of the POZ (POxvirus and Zinc finger) domain of the transcription factor Miz1 (Myc-interacting zinc finger protein;...

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