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The kinetics of the reaction require some optimization. The optimal incubation time for the reaction can vary depending on concentrations and parameters used and should be optimized by the end users in order for them to get the best window possible. Depending on the temperature and the length of the reactions, as well as the amounts of activator used, this window can vary. We normally advise customers to leave the plate and the 2X substrate solution at 37°C prior to initiating the reaction and then run a time course, which includes several time points over a period of 1-2 hours. Titration of resveratrol should also be envisaged.