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.
H3K4ac Antibody - replaced by the reference C15410322
(pAb-165-050)
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.
| Lot | A482-0042 |
|---|---|
| Concentration | 1.58 µg/µl |
| Species reactivity | Human, yeast |
| Type | Polyclonal |
| Purity | Affinity purified |
| Host | Rabbit |
| Precautions | This 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
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).
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.
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.
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.
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. - Publications
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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... - Related products
