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Assay Method Information

Assay Name:  Enzyme Inhibition Assay
Description:  Thrombin-catalyzed hydrolysis rates were measured spectrophotometrically using human alpha-thrombin, a chromogenic substrate in aqueous buffer, and a microplate reader. Changes in absorbance at 405 nm were monitored upon addition of enzyme both with and without inhibitor present. The IC50 values were determined by fixing the enzyme and substrate concentrations and varying the inhibitor concentration. Ki values were determined by fixing the enzyme and inhibitor concentrations and varying the substrate concentration. Michaelis-Menton kinetics was applied to the initial reaction slopes using the program K.Cat (Bio Metallics Inc.).
Affinity data for this assay
 

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Last update November 1, 2007
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