GSM3593238: H3K27ac_teloHAEC_24h_r1; Homo sapiens; ChIP-Seq
Cells were washed with HBSS Gibco and fixed in paraformaldehyde (PFA) 1% for 10 minutes at room temperature (RT). PFA was quenched in 134 mM glycine for 5 minutes at RT. Fixed cells were washed with ice-cold PBS and collected with a cell scraper in ice-cold PBS. Cells were pelleted by centrifugation, washed in ice-cold PBS, and pelleted again before snap freezing in liquid nitrogen. Fixed cells were subject to lysis in 5 mM PIPES-pH 8.5, 85 mM KCl, 1% (v/v) IGEPAL CA-630, 50 mM NaF, 1 mM PMSF, 1 mM Phenylarsine Oxide, 5 mM Sodium Orthovanadate and protease inhibitor cocktail. Nuclei were then lysed in 50 mM Tris-HCl pH 8.0, 10 mM EDTA, 1% (w/v) SDS, 50 mM NaF, 1 mM PMSF, 1 mM Phenylarsine Oxide, 5 mM Sodium Orthovanadate and protease inhibitor cocktail. Chromatin immunoprecipitation was performed as previously described using 3.7 μg of H3K27ac antibody (Diagenode) per samples containing 1 to 1.5 million of cells (no antibody for input). Library construction was performed using KAPA Hyper Prep Kit (Kapa Biosystems) for Illumina using with adapters from TruSeq DNA LT Sample Prep Kit (Illumina)
GEO Accession
GSM3593238