GSM7729125: 92-1,DMSO,4h,ChIP-seq,MITF; Homo sapiens; ChIP-Seq
Treated 92-1 cells were fixed with 1/10 volume freshly prepared Formaldehyde solution (37% Formaldehyde [Sigma], 5M NaCl [Sigma], 0.5 M EDTA pH 8.0 [Sigma], 1M HEPES pH 7.9 [Teknova]), and neutralized with 1/20 volumes of 2.5 M glycine (Sigma).  Cells were harvested and washed twice with ice-cold PBS (ThermoFisher) containing 0.5% Igepal (Sigma).  During the final wash, 1 mM PMSF was included. Cells were pelleted and following nuclei extraction, chromatin was sonicated. After centrifugating the sonicated chromatin for 10 min at 10,000 rpm, soluble fractions were incubated with the following antibodies: SMARCA4 (Abcam cat# ab110641), SOX10 (Abcam cat# ab155279), MITF (Active Motif cat# 39789), TFAP2A (Santa Cruz cat# sc-12726), H3K27ac (Active Motif cat# 39133). Illumina libraries were prepared by Active Motif using standard consecutive enzymatic steps of end-polishing, dA-addition, and adaptor ligation according to the standard Illumina ChIP-seq library.