| Assay Method Information | |
| | Enzyme Inhibition Assay |
| Description: | Enzyme assay was performed at room temperature on a microplate reader (Safire2TM) using black 96-well microtiter plates purchased from Nunc. Inhibitors and substrates were previously dissolved in 4% DMSO. The hydrolysis of the substrates was recorded as increase in fluorescence intensity. IC50 values were generated by nonlinear regression analysis from plots of vi/v0 versus inhibitor concentration, in which vi is the velocity in presence, and v0 the velocity in the absence of an inhibitor. The kinetic constants were determined by the method of Lineweaver and Burk, and Ki values were consecutively calculated from the following equation: Ki = IC50/[1 + (S/Km)]. The overall error of the assays is estimated to be +/-40%. |
| Affinity data for this assay | |
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