ChIP Analysis of H3K27me Polyclonal Antibody
ChIP assays were performed using human HeLa cells, the antibody against H3K27me3 (Cat. No. A15024) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP” kit (Cat. No. 4487119), using sheared chromatin from 1 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. QPCR was performed with primers for the promoters of the active genes EIF4A2 and GAPDH as negative controls, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. This figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3 is preferably present at inactive genes.