p300 monoclonal antibody - Classic

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against p300

ChIP was performed using HeLa cells, the Diagenode monoclonal antibody against p300 (cat. No. C15200211) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010055), using sheared chromatin from 4 million 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. Quantitative PCR was performed with primers for two genomic regions near the ANKRD32 and IRS2 genes, used as positive controls, and for the coding region of the inactive MYOD1 gene and an intergeic region on chromosome 11, 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 p300

ChIP was performed with 5 μg of the Diagenode antibody against p300 (cat. No. C15200211) on sheared chromatin from 4 million HeLa cells 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 3 mb region of chromosome 5 (figure 2A and B) and in two regions surrounding the IRS2 and ANKRD32 positive control genes (figure 2C and D). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.