Western blot analysis of extracts from various cell lines using ATRX (D1N2E) Rabbit mAb (upper) and Nucleolin (D4C7O) Rabbit mAb #14574. As expected, the signal for ATRX is not present in the negative cell line U2OS.
Immunoprecipitation of ATRX from 293T cell extracts using Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900 (lane 2) or ATRX (D1N2E) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ATRX (D1N2E) Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HeLa cells and either 10 µl of ATRX (D1N2E) 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 human 18s rDNA repeat primers, SimpleChIP ® Human 28s rDNA Repeat Primers #14901, and SimpleChIP ® Human GAPDH Exon 1 Primers #5516. 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.