Anti-SAP30 antibody (ab231804) at 1/1000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear extracts at 20 µg

Dilution buffer: TBS-Tween containing 5% skimmed milk. 

ChIP assays were performed using HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, ab231804 and optimized primer sets for qPCR. ChIP was performed using sheared chromatin from 4 million cells. A titration of the antibody consisting of 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 the EIF2S3 and BRCA1 promoters, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Image shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ChIP was performed on sheared chromatin from 4 million HeLa (Human epithelial cell line from cervix adenocarcinoma) cells using 5 µg ab231804. 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 50 bp tags were aligned to the human genome using the BWA algorithm. Image shows the enrichment along the complete sequence and a 1.5 Mb region of human chromosome 1 (A and B) and in two genomic regions surrounding the BRCA1 and EIF2S3 genes on chromosome 17 and X, respectively (C and D).