GSM6421759: Ishikawa_1hrE2_rep2; Homo sapiens; ChIP-Seq
Cells were fixed using 1% formaldehyde (Sigma Aldrich, 252549) for 10 minutes at room temperature to cross-link DNA and proteins. Cross-linking was stopped by addition of glycine (Fisher Scientific, BP381) to a final concentration of 125 mM. Cells were then washed with cold PBS and harvested via scraping in Farnham lysis buffer supplemented with protease and phosphatase inhibitors (Thermo Scientific, A32959). Chromatin immunoprecipitation was performed as previously described (Reddy et al. Genome Research 2009) with an Anti-Estrogen receptor alpha (ESR1) antibody (Millipore Sigma, 06-935) or H3K27ac antibody (Active Motif, 39133).