Anti-Rub1 Antibody - Western Blot. Anti-Rub1 antibody, generated by immunization with full-length, recombinant yeast Rub1, was tested by western blot against a yeast cell lysate. A dilution of the antibody between 1:200 and 1:1000 will show strong reactivity specifically with free Rub1 protein (indicated by arrow) and Rub1 conjugates. In this blot, the antibody was used at a 1:500 dilution and was incubated overnight at 4° C in 5% non-fat dry milk in TTBS. Detection occurred using a 1:2000 dilution of HRP-labeled Donkey anti-Rabbit IgG (code # LS-C60943) for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche).