GSM8621935: IRF2BP2; Homo sapiens; ChIP-Seq
Sonicated input sample (10% of total chromatin) was saved and subjected to crosslink reversing and column purification. Sonicated chromatin samples were incubated overnight at 4°C with IRF2BP2 antibody in 1 ml ChIP buffer (50 mM HEPES pH 7.8, 140 mM NaCl, 1% Triton X-100, 0.1% Na-Deoxycholate and Complete protease inhibitor). IP was washed and then eluted by incubation at 65°C overnight in 50 μl ChIP elution buffer (50 mM Tris pH 7.5, 10 mM EDTA, 1% SDS) with 10 mM DTT. Beads were pelleted and the supernatant was transferred to a new tube. DNA was subsequently isolated by columns (NucleoSpin, Takara). For ChIP-seq, ∼10 ng of ChIP DNA was prepared for sequencing according to the amplification protocol from Illumina.  ChIP-Seq libraries were prepared from formaldehyde-crosslinked hMSCs, using approximately 2–8 million cells per condition, depending on the abundance of the factor of interest.