Assay Method Information

Assay Name:  Enzyme Inhibition Assay
Description:  The enzyme activities were determined kinetically as the initial rate of cleavage of a peptide p-nitroanilide. Km for enzyme and substrate was determined using four substrate dilutions. Enzyme was incubated with test compounds for 10 min; the reaction was then started by the addition of the substrate. Reaction rates were determined by measuring the rate of the absorbance change at 405 nm in a ThermoMax microplate reader (Molecular Devices Corp., Sunnyvale, CA). IC50 values for inhibitors were determined from the dose response curve by fitting the data to the Hill equation. Ki,app values were calculated as the IC50/2, with the IC50 value determined at a substrate concentration equal to Km
Affinity data for this assay
 

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