Fig. 3
From: Robust high-throughput assays to assess discrete steps in ubiquitination and related cascades
Alpha assays for Uba1~ubiquitin thioester formation, Rad6~ubiquitin thioester formation, and Rad18 ubiquitination. Data represent mean and SD for 5 samples in each case. Variable concentrations of a FLAG-Uba1 and b biotin-ubiquitin (Bio-Ub) in the Uba1~ubiquitin thioester formation assay, with the other component in each case being held constant. Variable concentrations of c FLAG-Rad6 and d biotin-ubiquitin in the Rad6~ubiquitin thioester formation assay. Variable concentrations of e FLAG-Rad18 and f biotin-ubiquitin in the Rad18 autoubiquitination assay. Details for each assay are described in Methods