GSM2650785: Input control sample; Homo sapiens; ChIP-Seq
U937 cells stablely expressing Flag-PtpA were seeded at 1×107 cells per 10 cm dish. Cells were crosslinked with 1% formaldehyde for 10 min. Glycine was then added for 5 min to a final concentration of 0.125 M, cells scrapped and collected. The pellet was resuspended in lysis buffer (5 mM Tris-HCl, pH 8.0, 85 mM KCl, 0.5% NP40 supplemented with protease inhibitors cocktail) and subjected to centrifugation to collect nuclear pellet. The pellet was resuspended in RIPA buffer (20 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM EDTA, 0.5% NP40, 0.5% Na-deoxycholate, 0.1% SDS supplemented with protease inhibitors cocktail). The nuclear extracts were sonicated. The chromatin was incubated overnight with anti-Flag antibody (Sigma) followed by four washes with LiCl wash buffer (100 mM Tris-HCl, pH 7.5, 500 mM LiCl, 1% NP40, 1% Na-deoxycholate) and 1 wash with TE (10 mM Tris-HCl, pH 7.5, 1 mM EDTA, pH 8.0). DNA was eluted in Elution Buffer (1% SDS, 0.1 M NaHCO3) for 1 h at 37℃ and used for sequencing library preparation.
GEO Accession
GSM2650785