Diagenode

H3K36ac polyclonal antibody - Classic (sample size)

Histone-Deacetylase-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410307-10
10 μg
$80.00
  Bulk order
Other format

Polyclonal antibody raised in rabbit against the region of histone H3 containing the acetylated lysine 36 (H3K36ac), using a KLH-conjugated synthetic peptide.

LotA2238P
Concentration0.9 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 * 2 μg/ChIP Fig 1, 2
ELISA 1:1,000 Fig 3
Dot Blotting 1:10,000 Fig 4
Western Blotting 1:1,000 Fig 5

* 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

    ChIP

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K36ac
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K36ac (cat. No. C15410307) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010055), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the ACTB promoter and for the GAPDH promoter, used as positive controls, and for the coding region of 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).

    ChIP-seq results A

    ChIP-seq results B

    ChIP-seq results C

    ChIP-seq results D

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K36ac
    ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K36ac (cat. No. C15410307) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. 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 enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 genes.

    ELISA

    Figure 3. 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 H3K36ac (cat. No. C15410307) in antigen coated wells. 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 1:10,000.

    Cross reactivity

    Figure 4. Cross reactivity tests using the Diagenode antibody directed against H3K36ac
    To test the cross reactivity of the Diagenode antibody against H3K36ac (cat. No. C15410307), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K36. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

    Western blot

    Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36ac
    Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K36ac (cat. No. C15410307). The antibody was 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.

  • 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-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  • Documents
    Datasheet H3K36ac C15410307 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...
    Download
  • Publications

    How to properly cite this product in your work

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Events

  • Workshop on Chromatin Proteomics
    Crete, Greece
    Oct 3-Oct 8, 2016
  • 2nd Annual Next Generation Sequencing Congress
    Boston, USA
    Oct 3-Oct 4, 2016
  • XXXIX Reunión anual de la Sociedad de Bioquimica y Biología Molecular de Chile
    Puerto Varas, Chile
    Sep 27-Sep 30, 2016
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