Anti-Brd4 (phospho Thr204) Antibody [ARC50960] (A305446)

Western blot analysis of extracts of various cell lines, using Anti-Brd4 (phospho Thr204) Antibody [ARC50960] (A305446) at 1:1,000 dilution. HeLa cells were treated by Calyculin A (100 nM) at 37°C for 30 minutes after serum-starvation overnight. C6 cells were treated by Calyculin A (100 nM) at 37°C for 30 minutes after serum-starvation overnight. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% non-fat dry milk in TBST. Detection was with a ECL Basic Kit. Exposure time: 1s.

Western blot analysis of extracts of NIH/3T3 cells, using Anti-Brd4 (phospho Thr204) Antibody [ARC50960] (A305446) at 1:1,000 dilution. NIH/3T3 cells were treated by Calyculin A (100 nM) for 30 minutes with NaCl (400mM) after serum-starvation 16-20 hours. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% non-fat dry milk in TBST. Detection was with a ECL Enhanced Kit (RM00021). Exposure time: 90s.