Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and either 5 μl of SRC-3 (5E11) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP ® Human CyclinD1 Promoter Primers #12531, human CTSD promoter primers, and SimpleChIP ® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Western blot analysis of cell lysates from A549, HeLa and Jurkat cells using SRC-3 (5E11) Rabbit mAb.