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 10 μl of Oct-1 (D7B6) 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 ESR1 Promoter Primers #9673, SimpleChIP ® Human pS2 Promoter Primers #9702, 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.
Westerrn blot analysis of extracts from various cell lines using Oct-1 (D7B6) Rabbit mAb.