Immunoprecipitation of SS18 from ACHN cell extracts. Lane 1 is 10% input, lane 2 is Rabbit mAb IgG Isotype control, and lane 3 is SS18 Rabbit mAb. Western blot analysis was performed using SS18 Rabbit mAb.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4x10e6 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with beta-estradiol (10nM) for 45 min and either 10ul of SS18 Rabbit mAb or 2ul of Normal Rabbit IgG using Enzymatic Chromatin IP Kit (Magnetic Beads). The enriched DNA was quantified by real-time PCR using Human ESR1 Promoter Primers Human pS2 Promoter Primers, and Human alpha Satellite Repeat Primers. 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.