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Moreover, the p53 was phosphorylated, translocated into nucleus, and bound to Rp following ROS stimulation

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The unimolecular approach is simpler but less flexible

In the reaction mixtures without enzyme, varying amounts of MBB (0-300 l of 1 mM stock solution which gave final concentration 0-100 M) were added into different fluorescent cuvettes already containing 3.0 ml of phosphate buffer (pH 6.5) and 100 l of GSH stock solution (3 mM)