H3K4ac Antibody - replaced by the reference C15410322

Catalog Number
50 μg/34 μl

As an alternative we offer H3K4ac polyclonal antibody - Classic (C15410322)

Polyclonal antibody raised in rabbit against histone H3 acetylated at lysine 4 (H3K4ac), using a KLH-conjugated synthetic peptide.

Concentration1.58 µg/µl
Species reactivityHuman, yeast
PurityAffinity purified
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 0.5 μg/ChIP Fig 1
ELISA 1:500 Fig 2
Dot Blotting 1:10,000 Fig 3
Western Blotting 1:500 Fig 4
Immunofluorescence 1:300 Fig 5

* Please note that the optimal antibody amount per IP should be determined by the end-user.

  • Validation Data

    H3K4ac Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac
    ChIP assays were performed using WT and H3K4R mutant S. pombe cells, the Diagenode antibody against H3K4ac (Cat. No. C15410165) and optimized primer pairs for qPCR. Sheared chromatin corresponding to 10 μg of DNA and 0.5 μg of antibody were used per ChIP experiment. QPCR was performed using primers specific for two different pericentric repeat regions and for the euchromatic adh1 gene. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA).

    H3K4ac Antibody ELISA validation

    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 H3K4ac (Cat. No. C15410165) 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 2), the titer of the purified antibody was estimated to be 1:27,800.

    H3K4ac Antibody validated in Dot blot

    Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K4ac
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410165) with peptides containing other histone H3 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 3 shows a high specificity of the antibody for the modification of interest.

    H3K4ac Antibody validated in Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K4ac
    Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode antibody directed against H3K4ac (Cat. No. C15410165), diluted 1:500 in TBS-Tween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.

    H3K4ac Antibody validated in Immunofluorescence

    Figure 5. Immunofluorescence with the Diagenode antibody directed against H3K4ac
    Wild type and H3K4R mutant S. pombe cells were stained with both the Diagenode antibody against H3K4ac (Cat. No. C15410165) (in red) and by Hoechst staining (in blue, left), or with the H3K4ac antibody alone (right). The antibody was used at a dilution of 1:300.

  • 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
    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 H3K4ac pAb-165-050 DATASHEET
    Polyclonal antibody raised in rabbit against histone H3 acetylated at lysine 4 (H3K4ac), using a ...
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
  •  Publications

    How to properly cite this product in your work

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    TIP60: an actor in acetylation of H3K4 and tumor development in breast cancer.
    Judes G, Dubois L, Rifaï K, Idrissou M, Mishellany F, Pajon A, Besse S, Daures M, Degoul F, Bignon YJ, Penault-Llorca F, Bernard-Gallon D
    AIM: The acetyltransferase TIP60 is reported to be downregulated in several cancers, in particular breast cancer, but the molecular mechanisms resulting from its alteration are still unclear. MATERIALS & METHODS: In breast tumors, H3K4ac enrichment and its link with TIP60 were evaluated by chromatin immunoprecip...

    H3K4 acetylation, H3K9 acetylation and H3K27 methylation in breast tumor molecular subtypes
    Judes G et al.
    AIM: Here, we investigated how the St Gallen breast molecular subtypes displayed distinct histone H3 profiles. PATIENTS & METHODS: 192 breast tumors divided into five St Gallen molecular subtypes (luminal A, luminal B HER2-, luminal B HER2+, HER2+ and basal-like) were evaluated for their histone H3 modifica...

    Epigenetic Modifications with DZNep, NaBu and SAHA in Luminal and Mesenchymal-like Breast Cancer Subtype Cells
    Dagdemir A et al.
    BACKGROUND/AIM: Numerous studies have shown that breast cancer and epigenetic mechanisms have a very powerful interactive relation. The MCF7 cell line, representative of luminal subtype and the MDA-MB 231 cell line representative of mesenchymal-like subtype were treated respectively with a Histone Methyl Transferas...

    Molecular and Epigenetic Biomarkers in Luminal Androgen Receptor: A Triple Negative Breast Cancer Subtype
    Judes G et al.

    Histone lysine trimethylation or acetylation can be modulated by phytoestrogen, estrogen or anti-HDAC in breast cancer cell lines.
    Dagdemir A, Durif J, Ngollo M, Bignon YJ, Bernard-Gallon D
    AIM: The isoflavones genistein, daidzein and equol (daidzein metabolite) have been reported to interact with epigenetic modifications, specifically hypermethylation of tumor suppressor genes. The objective of this study was to analyze and understand the mechanisms by which phytoestrogens act on chromatin in breast c...

    A chromodomain switch mediated by histone H3 Lys 4 acetylation regulates heterochromatin assembly.
    Xhemalce B, Kouzarides T
    Chromodomain proteins (Chp1/Chp2/Swi6/Clr4) bind to methylated H3K9 (H3K9me) and regulate pericentric heterochromatin in fission yeast. Chp1 and Clr4 (H3K9-HMT), bind transcriptionally active heterochromatin, whereas Chp2/Swi6 (HP1 homologs) are recruited during the inactive state. We show that H3K4 acetylation (H3K...

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