Anti-Ubiquitin antibody, generated by immunization with Ubiquitin coupled to Rabbit IgG, was tested by WB against total cell extract from yeast. Dilution of the antibody between 1:200 and 1:1,000 showed strong reactivity with Ubiquitinated proteins. In this blot the antibody was used at 1:500. Detection occurred using a 1:2000 dilution of HRP-labeled Donkey anti-Rabbit IgG.