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| | 3alpha-Androstanediol Oxidation Inhibition Assay |
| Description: | A fluorescence assay was used to determine the kinetic constants for the oxidation of 3alpha-androstanediol. The fluorescence emission of NADPH at 450 nm (5-nm slit width) with excitation at 340 nm (15-nm slit width) was monitored. A standard curve was constructed by monitoring fluorescence changes with incremental additions of NADPH. For inhibition studies, IMN and FLF were added at several different concentrations to give final concentrations varying between 0.5 and 50 uM, and steroid substrate concentrations were then varied as indicated above. The type of inhibition and the inhibition constants were calculated using the Leonora program. All reactions were initiated by the addition of enzyme. |
| Affinity data for this assay | |
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