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10 U of thrombin or 100 µg of native and deglycosylated rhinocerase were mixed with different concentrations of PMSF and incubated at 37°C for 30 minutes prior to the addition of Arg-AMC to a final concentration of 50 nM. The amount of Arg-AMC released was measured after 15 minutes of incubation using a spectrofluorometer. Each bar shows the mean ± S.D. (n = 3). The hydrolytic activity measured for rhinocerase without any PMSF was taken as 100%.
