N-[5-Carbamimidamido-1-(4-nitroanilino)-1-oxopentan-2-yl]-1-phenylalanylpiperidine-2-carboximidic acid → 4-Nitroaniline

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Reaction Steps

An inhibitor sample is dissolved in a 0.05 M Tris-hydrochloric acid buffer solution (pH=8.3) to make a total volume of 400 μl. To this solution, 50 μl of a 0.2 mM S-2238 solution is added and the mixture is incubated at a temperature of 37° C. for 5 minutes in a constant temperature bath. Then, 50 μl of a 0.2 unit/ml bovine thrombin solution is added thereto and the absorbance, at 405 nm, of the p-nitroaniline formed per minute is determined at a temperature of 37° C. by using the so-called initial velocity method. Thus, the concentration I50 (μmol) of the inhibitor sample at which the absorbance is one half (i.e., 1/2) of that obtained in the absence of the inhibitor sample, is determined.

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