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In vitro Protein Degradation Assays
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20 nM CP alone or CP reconstituted with 60 nM RP was incubated with ~100 nM substrate, 1 mM ATP, 0.5 mM DTT, and 0.4 mg/ml BSA. At each time point, 10 μl of the reaction mixture was withdrawn, and the reaction was terminated by adding Laemmli loading buffer. Samples were then analyzed by SDS-PAGE and immunoblotting.
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