Assay Method Information |
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| Determination of Dissociation Constant (Kd) |
Description: | Values of Ki were determined from the dependence of the observed rate constant (kobs) on inhibitor concentration. With subsaturing APS, the inhibition constant is equal to the dissociation constant (Ki = Kd). A range of inhibitor concentrations was employed from at least 5-fold below to 10-fold above the inhibition constant. Nonlinear least-squares fits of the equation for competitive inhibition gave excellent fits in all cases, and the standard error was typically less than 15%. |
Affinity data for this assay | |
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