Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 MCF7 cells grown in phenol red free medium and charcoal stripped FBS for 4 d and then treated with β-estradiol (10 nM) for 1 hr and either 5 μl of SIN3A (D1B7) 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 GAPDH Exon 1 Primers #5516, SimpleChIP ® Human MyoD1 Exon 1 Primers #4490, 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 extracts from various cell lines using SIN3A (D1B7) Rabbit mAb.