Diagenode

H3K36me1 polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410089
(pAb-089-050)
50 µg/54 µl
$295.00
  Bulk order

Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 36 (H3K36me1), using a KLH-conjugated synthetic peptide.

LotA2367-001P
Concentration0.94 µg/µl
Species reactivityHuman
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 5 μg/ChIP Fig 1
ELISA 1:100 Fig 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:1,000 Fig 4
Immunofluorescence 1:500 Fig 5
* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
  • Validation Data

    ChIP

    Figure 1 ChIP results obtained with the Diagenode antibody directed against H3K36me1
    ChIP assays were performed using human U2OS cells, the Diagenode antibody against H3K36me1 (cat. No. pAb-089-050) and optimized PCR primer pairs for qPCR. ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells and stringent washing conditions. A titration consisting of 1, 2, 5 and 10 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 for the promoter of the constitutively expressed GAPDH gene and for the inactive 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).

    ELISA

    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 H3K36me1 (cat. No. pAb-089-050), crude serum and Flow Through. 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 1:46,000.

    Dot Blot

    Figure 3 Cross reactivity tests using the Diagenode antibody directed against H3K36me1
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me1 (cat. No. pAb-089-050) with peptides containing other modifications of histone H3 or the unmodified H3K36. 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 1:20,000. 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 H3K36me1
    Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me1 (cat. No. pAb-089-050) 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.

    Immunofluorescence

    Figure 5 Immunofluorescence using the Diagenode antibody directed against H3K36me1
    Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K36me1 (cat. No. pAb- 089-050) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K36me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K36me1 antibody after incubation of the antibody with 2 ng/μl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K36, respectively.

  • 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
    ChIP-qPCR (ab)
    Read more
  • Documents
    Datasheet H3K36me1 pAb-089-050 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...
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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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  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K36me1 polyclonal antibody - Classic (Diagenode Cat# C15410089 Lot# A2367-001P). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Chromatin states of core pluripotency-associated genes in pluripotent, multipotent and differentiated cells.
    Barrand S, Collas P
    Oct4, Nanog and Sox2 constitute a core of transcription factors controlling pluripotency. Differentiation and reprogramming studies have unraveled a few epigenetic modifications associated in relation to the expression state of OCT4, NANOG and SOX2. There is, however, no comprehensive map of chromatin states on thes...

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