Figure. Immunoblot of Rub1 fusion protein. Anti-Rub1 antibody, generated by immunization with full-length recombinant yeast Rub1, was tested by immunoblot against a yeast cell lysate. A dilution of the antibody between 1:200 and 1:1,000 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 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 for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche). Other detection systems will yield similar results.
