H3K9ac Antibody

Catalog Number
50 μg

As an alternative we offer the purified H3K9ac polyclonal antibody - Classic (C15410004) 

Polyclonal antibody raised in rabbit against histone H3, acetylated at lysine 9 (H3K9ac), using a KLH-conjugated synthetic peptide.

Concentration1.08 µg/µl
Species reactivityHuman
PurityAffinity purified
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1-2 μg/ChIP Fig 1, 2
ELISA 1:1,000 Fig 3
Dot Blotting 1:20,000 Fig 4
Western Blotting 1:750 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

    H3K9ac Antibody ChIP-seq Grade

    H3K9ac Antibody for ChIP-seq

    H3K9ac Antibody for ChIP-seq assay

    H3K9ac Antibody validated in ChIP-seq

    Figure 1 ChIP-seq results obtained with the Diagenode antibody directed against H3K9ac
    ChIP was performed as described above using 1 μg of the Diagenode antibody against H3K9ac (cat. No. pAb- 177-050). The IP’d DNA was subsequently analysed with 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 peak distribution along the complete sequence and a 800 kb region of the X-chromosome (figure 2A and B) and in 100 kb regions surrounding the GAPDH and c-fos genes (figure 2C and D). These results clearly show an enrichment of H3K9ac at the promoters of active genes.

    H3K9ac Antibody ChIP Grade

    Figure 2 ChIP results obtained with the Diagenode antibody directed against RARA
    ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K9ac (cat. No. pAb-177-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (cat. No. AB- Auto02-A100), using sheared chromatin from 1 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. Quantitative PCR was performed with primers for the promoter of the active c-fos (cat. No. pp-1004-050) and GAPDH genes, used as positive controls, and for the coding region of the inactive MB gene (cat. No. pp-1006-050) 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). These results are in accordance with the observation that H3K9 acetylation is enriched at the promoters of active genes.

    H3K9ac Antibody ELISA validation

    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 H3K9ac (cat. No. pAb-177-050) 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 1:59,600.

    H3K9ac Antibody validated in Dot Blot

    Figure 4 Cross reactivity test using the Diagenode antibody directed against H3K9ac
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9ac (cat. No. pAb-177-050) with peptides containing other histone modifications and the unmodified H3K9 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 4 shows a high specificity of the antibody for the modification of interest. Please note that this antibody recognizes the H3K9 acetylation, both in the presence and the absence of the K14 acetylation or the S10 phosphorylation.

    H3K9ac Antibody validated in Western Blot

    Figure 5 Western blot analysis using the Diagenode antibody directed against RARA
    Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9ac (cat. No. pAb-177-050) 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.

  • 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
    Enzyme-linked immunosorbent assay. Read more
    Dot blotting Read more
    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 H3K9ac pAb-177-050 DATASHEET
    Datasheet description
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K9ac Antibody (Diagenode Cat# C15410177 Lot# A1434-0012P). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    MeCP2 regulates gene expression through recognition of H3K27me3.
    Lee, W and Kim, J and Yun, JM and Ohn, T and Gong, Q
    MeCP2 plays a multifaceted role in gene expression regulation and chromatin organization. Interaction between MeCP2 and methylated DNA in the regulation of gene expression is well established. However, the widespread distribution of MeCP2 suggests it has additional interactions with chromatin. Here we demonstra...

    Remodelling of the hepatic epigenetic landscape of glucose-intolerant rainbow trout (Oncorhynchus mykiss) by nutritional status and dietary carbohydrates.
    Marandel L et al.
    The rainbow trout, a carnivorous fish, displays a ‘glucose-intolerant’ phenotype revealed by persistent hyperglycaemia when fed a high carbohydrate diet (HighCHO). Epigenetics refers to heritable changes in gene activity and is closely related to environmental changes and thus to metabolism adjustments g...

    Epigenetic silencing of serine protease HTRA1 drives polyploidy
    Schmidt N et al.
    BACKGROUND: Increased numbers and improperly positioned centrosomes, aneuploidy or polyploidy, and chromosomal instability are frequently observed characteristics of cancer cells. While some aspects of these events and the checkpoint mechanisms are well studied, not all players have yet been identified. As the ro...

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