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H3K9acS10p Antibody - ChIP Grade

Catalog Number
Format
Price
C15310102
(CS-102-100)
100 µl
$332.00
  Bulk order
Other format



Polyclonal antibody raised in rabbit against histone H3 containing the acetylated lysine 9 and the phosphorylated serine 10 (H3K9acS10p), using a KLH-conjugated synthetic peptide.

LotA397-001
Concentrationnot determined
Species reactivityHuman
TypePolyclonal
PurityWhole antiserum
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 15 μl/ChIP Fig 1
ELISA 1:1,000 - 1:4,000 Fig 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:250 Fig 4
IF 1:500 Fig 5
* 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

    H3K9acS10p Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p
    ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K9acS10p Antibody ELISA validation

    Figure 2. Determination of the titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.

    H3K9acS10p Antibody validated in Dot Blot

    Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. 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 double modification.

    H3K9acS10p Antibody validated in Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p
    Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.

    H3K9acS10p Antibody validated in Immunofluorescence

    Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p
    HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) 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 H3K9acS10p 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. Acetylation of K9 and phosphorylation of S10 of histone H3 are associated with active gene transcription

  •  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-qPCR (ab)
    Read more
  •  Documents
    Datasheet H3K9acS10p CS-102-100 DATASHEET
    Datasheet description
    Download
  •  Publications

    How to properly cite this product in your work

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