Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HCT 116 cells starved for 48 hr then serum stimulated with 20% FBS for 15 min and either 10 μl of SSRP1 (E1Y8D) 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 c-Fos Promoter Primers #4663, SimpleChIP ® Human c-Fos Exon 3 Primers #12010, SimpleChIP ® Human EGR1 Promoter Primers #5549, and SimpleChIP ® Human EGR1 Intron 3 Primers #11953. 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 SSRP1 (E1Y8D) Rabbit mAb.