H3K27ac monoclonal antibody

50 μg
  Bulk order

Monoclonal antibody raised in mouse against histone H3 acetylated at lysine 27 (H3K27ac), using a KLH-conjugated synthetic peptide.

Concentration1.0 µg/µl
Species reactivityHuman, Nematodes: positive. Other species: not tested.
PurityProtein A purified
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 * 1-2 μg/ChIP Fig 1
CUT&TAG 1:3,000 Fig 2
ELISA 1:3,000 Fig 3
Western Blotting 1:1,000 - 1:2,000
Immunofluorescence 1:500 Fig 4

* 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

    H3K27ac Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27ac
    ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K27ac (Cat. No. C15200184) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010020), using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the EIF4A2 and GAPDH genes, used as positive controls, and for 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).



    Figure 2. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K27ac
    CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K27ac (cat. No. C15200184) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence of chromosome 7 and in a 250 kb region surrounding the ESWR1 gene on chromosome 12 (figure 2A and B, respectively).

    H3K27ac Antibody ELISA validation

    Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against H3K27ac
    To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K27ac (Cat. No. C15410184). The wells were coated with peptides containing the unmodified H3K27 region as well as the acetylated H3K27 and the acetylated H3K9. Figure 2 shows a high specificity of the antibody for the peptide containing the modification of interest.

    H3K27ac Antibody validated in Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K27ac
    HeLa cells were stained with the Diagenode antibody against H3K27ac (Cat. No. C15410184) 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 H3K27ac antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. 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.

  •  应用
    Enzyme-linked immunosorbent assay. 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
    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
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  •  文档
    Datasheet H3K27ac MAb-184-050 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...
  •  Safety sheets
    H3K27ac antibody SDS GB en Download
    H3K27ac antibody SDS US en Download
    H3K27ac antibody SDS BE nl Download
    H3K27ac antibody SDS BE fr Download
    H3K27ac antibody SDS FR fr Download
    H3K27ac antibody SDS ES es Download
    H3K27ac antibody SDS DE de Download
    H3K27ac antibody SDS JP ja Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K27ac monoclonal antibody (Diagenode Cat# C15200184-50 Lot# 001-15). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Systematic discovery and functional dissection of enhancers needed forcancer cell fitness and proliferation.
    Chen Poshen B et al.
    A scarcity of functionally validated enhancers in the human genome presents a significant hurdle to understanding how these cis-regulatory elements contribute to human diseases. We carry out highly multiplexed CRISPR-based perturbation and sequencing to identify enhancers required for cell proliferation and fitness ...

    Evidence for long-term association of virion-delivered HBV core proteinwith cccDNA independently of viral protein production
    Lucifora Julie et al.
    Background \& Aims HBV persists in the nucleus of infected hepatocytes as a covalently closed circular DNA (cccDNA) episome that constitutes the template for viral RNA and protein synthesis. Both HBx and HBc (core) viral proteins associate with cccDNA but, while HBx is required for viral transcription, the role ...

    Potent and selective bivalent inhibitors of BET bromodomains
    Waring M.J. et al.
    Proteins of the bromodomain and extraterminal (BET) family, in particular bromodomain-containing protein 4 (BRD4), are of great interest as biological targets. BET proteins contain two separate bromodomains, and existing inhibitors bind to them monovalently. Here we describe the discovery and characterization of pro...

    Germline organization in Strongyloides nematodes reveals alternative differentiation and regulation mechanisms.
    Kulkarni A et al.
    Nematodes of the genus Strongyloides are important parasites of vertebrates including man. Currently, little is known about their germline organization or reproductive biology and how this influences their parasitic life strategies. Here, we analyze the structure of the germline in several Strongyloides and closely ...

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