| Assay Method Information | |
| | Cyclophilin Peptidyl-Prolyl Isomerase Functional Assay |
| Description: | Measurements were performed using an Agilent 8453 spectrophotometer. Assay buffer was cooled to 10° C. (with stirring) in a precision glass cuvette and inhibitor was added from a DMSO stock solution to afford a final concentration of <1% DMSO. A blank spectrum was obtained and then enzyme and substrate were added and the change in absorbance measured over 5 min. A first order rate was fitted to the absorbance data to obtain a rate constant (first 10 to 15 s were eliminated due to mixing). The catalytic rate was calculated from the enzymatic rate minus the background rate. The enzymatic rate constant, determined in duplicate at each inhibitor concentration, was plotted against inhibitor concentration and a non-linear fit by SigmaPlot generated a Ki. |
| Affinity data for this assay | |
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