RUB1 polyclonal antibody (Cat # PAB11355), 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 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 for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche).
Ubiquitin-like protein with similarity to mammalian NEDD8; conjugation (neddylation) substrates include the cullins Cdc53p, Rtt101p, and Cul3p; activated by Ula1p and Uba3p (E1 enzyme pair); conjugation mediated by Ubc12p (E2 enzyme)