GSM2572277: PC3 (OE Pbx4)-Pbx4; Homo sapiens; ChIP-Seq
Cells were fixed with 1% formaldehyde for 10mins at room temperature, and stopped by adding glycine to a final concentration of 0.2M. Cells were washed with ice cold TBSE(20mM Tris-HCl, 150mM NaCl, 1mM EDTA) for three times. Chromatin were then extracted and sonicated to ~500bp. Input DNA was precleared with protein G Dynabeads (Life technologies), and followed by chromatin immunoprecipitation with Hoxc5 (HPA026794, Sigma) and PBX4 (HPA049859, Sigma) at 4°C overnight. ChIP beads were washed six times at room temperature. At least 10ng ChIP DNA were used to prepare libraries with NEBNext ChIP-Seq library prep reagent set (New England Biolabs), and multiplexed (New England Biolabs).