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. Author manuscript; available in PMC: 2012 May 1.
Published in final edited form as: Anal Biochem. 2011 Jul 2;418(1):102–110. doi: 10.1016/j.ab.2011.06.034

Figure 4. High throughput screen for the effect of ubiquitin mutations on polyubiquitin chain formation.

Figure 4

The effect of ubiquitin mutations on polyubiquitin chain formation by Ubc13-Mms2. Wells in row 1 contain E1, Ubc13-Mms2 and ubiquitin, row 2 contains E1 and ubiquitin, and row 3 contains ubiquitin alone. Protein concentrations used were 2 μM Ubc13-Mms2, 0.5 μM E1, 500 μM ubiquitin (except for no-lysine ubiquitin which was at 100 μM). Reactions were incubated for 1 hour at 37 °C, quenched and the absorbance measured. Assay was performed in duplicate, with representative data shown.