GSM5011994: hFOB-SMAD1-IP-with-BMP2-N1; Homo sapiens; ChIP-Seq
Cells were fixed in 1% formaldehyde and quenched by adding glycine 125 mM for 5 min at 4 °C. After this, cells were harvested and lysed according to the protocol described by Lee et al. and chromatin was sheared by sonication using the Bioruptor ultrasonicator (Diagenode). Sheared chromatin was immunoprecipitated using 10 μg of total SMAD1 antibody (Cell Signaling, #9743) or the normal rabbit IgG antibody (Cell Signaling, #2729) as a control, followed by incubation with protein G magnetic beads (Invitrogen). Eluted DNA was purified using DNA purification buffers and spin columns kit, according to the manufacturer's instructions (Cell Signaling). NEBNext Ultra II DNA library preparation kit was used according to manufacturer's instruction.