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.
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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.
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
| 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 H3K4ac pAb-165-050 DATASHEET Polyclonal antibody raised in rabbit against histone H3 acetylated at lysine 4 (H3K4ac), using a ... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
Publications
How to properly cite our product/service in your workWe strongly recommend using this: H3K4ac Antibody - replaced by the reference C15410322 (Hologic Diagenode Cat# C15410165 Lot# A482-0042). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
TIP60: an actor in acetylation of H3K4 and tumor development in breast cancer. |
H3K4 acetylation, H3K9 acetylation and H3K27 methylation in breast tumor molecular subtypes |
Epigenetic Modifications with DZNep, NaBu and SAHA in Luminal and Mesenchymal-like Breast Cancer Subtype Cells |
Molecular and Epigenetic Biomarkers in Luminal Androgen Receptor: A Triple Negative Breast Cancer Subtype |
Histone lysine trimethylation or acetylation can be modulated by phytoestrogen, estrogen or anti-HDAC in breast cancer cell lines. |
A chromodomain switch mediated by histone H3 Lys 4 acetylation regulates heterochromatin assembly. |