| Lot | 001 |
|---|---|
| Concentration | 1 µg/µl |
| Species reactivity | Human. Other species not tested. |
| Type | Monoclonal |
| Purity | Protein A purified monoclonal antibody in PBS containing 50% glycerol, 1% BSA and 0.09% azide. |
| Host | Rabbit |
| Storage Conditions | Store at -20°C |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 0.5 µg/ChIP | Fig 1, 2 |
| Western Blotting | 1:2,000 | Fig 3 |
| Immunofluorescence | 1:500 | Fig 4 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 μg per IP.

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H2A.ZK4ac
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2A.ZK4ac (cat. No. C15210009) and optimized PCR primer sets for qPCR. ChIP was performed with the "iDeal ChIP-seq" kit (cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the coding region of the active CCT5 gene and the Sat2 satellite repeat region 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. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H2A.ZK4ac
ChIP was performed with 0.5 µg of the Diagenode antibody against H2A.ZK4ac (cat. No. C15210009) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the EIF4A2 and GAPDH positive control genes (figure 2C and D).

Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H2A.ZK4ac
Whole cell extracts from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H2A.ZK4ac (cat. No. C15210009) diluted 1:2,000 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.

Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H2A.ZK4ac
HeLa cells treated with sodium butyrate were stained with the Diagenode antibody against H2A.ZK4ac No. C15210009, red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).
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 socalled "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of H2A.ZK4 is associated with gene activation.
| ChIP-seq (ab) Read more |
| ChIP-qPCR (ab) Read more |
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