H3K56ac polyclonal antibody

Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac
ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of 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).

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Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac
ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.

Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac
CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) 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 in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).

Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac
Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 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.