Fig. 2
From: Robust high-throughput assays to assess discrete steps in ubiquitination and related cascades
Alpha assay for PCNA ubiquitination. Data represent mean and standard deviation (SD) for 3–6 replicate samples in each case. Variable concentrations of: a FLAG-PCNA, b biotin-ubiquitin (Bio-Ub), c RFC (left and right panels are different concentration ranges from separate experiments), d Uba1, and e Rad6–Rad18 dimer, with other components in each case held constant as noted in Methods. f Reaction kinetics under final optimized conditions. Details are described in Methods