WB
Western blot analysis of lysates from PC-3 cells, using Phospho-Ubiquitin-Ser65 Rabbit pAb at 1:400 dilution. PC-3 cells were treated with Carbonyl cyanide 3-chlorophenylhydrazone(10 uM) at 37℃ for6 hours after serum-starvation overnight.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Enhanced Kit.Exposure time: 60s.