Western blot analysis of extracts from various cell lines using Estrogen Receptor α (D6R2W) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of MCF7 (ERα positive, left) and SK-BR-3 (ERα negative, right) cells using Estrogen Receptor α (D6R2W) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5 ® #4084 (fluorescent DNA dye).
Flow cytometric analysis of MD-MBA-231 (blue) and MCF7 cells (green) using Estrogen Receptor α (D6R2W) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragments (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
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 days, then treated with β-estradiol (10 nM) for 45 minutes and either 10 μl of Estrogen Receptor α (D6R2W) 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.