GSM8325847: HK2_IgG_2_Med; Homo sapiens; RIP-Seq
Cells were trypsinized and extracts were isolated with lysis buffer with RNase Inhibitor (40 U/μl; Thermo Scientific). Buffers included protease inhibitor cocktail (Sigma-Aldrich). Lysates were immunoprecipitated with Abs against FLAG or mouse IgG1. Beads were washed with NT2 buffer and digested with DNase I (Roche Applied Science) and protease K (Sigma-Aldrich). Total RNA was extracted with acid phenol or TRIzol. RIP-seq libraries were prepared using SMART-Seq v4 Ultra Low Input RNA kit