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A solution containing Fluor de Lys SIRT1 substrate (1 μM final concentration) and 10 mM KCl (final concentration) was added to 5 nM HDAC3:NCOR2 (final concentration) in 25 mM Tris pH 8 with 500 μM EDTA. Initial velocities were determined from fluorescence changes over time, normalized, plotted versus 10-HDA concentration, and fit using Eq 2.
