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Top1-Clamp activity is reversibly inhibited by diamide. Equal concentrations of Top1 and Top1-Clamp were incubated in plasmid DNA relaxation assays in the presence of 5 mM diamide and 100 mM DTT as indicated. Plasmid DNA was added at time 0, and the extent of DNA relaxation was assessed after 30 sec, 10 min, and 20 min at 37°C. Lane C, untreated plasmid DNA.
