The wild-type Baculo-RB, the wild-type GST-RB and the GST-RB-S780L in which Leu is substituted for Ser780of the wild-type pRB were incubated with or without (control) cyclin-Cdk in reaction buffer containing 100M cold ATP.
Then these RN proteins were subjected to SDS-PAGE and analyzed using Western blotting with the anti-phosphoRB (Ser780) and anti-RB antibody 3H9. Anti-RB antibody 3H9 reacts with both phosphorylated and unphosphorylated types of pRB.
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