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Assay Method Information

Assay Name:  Inhibition Study on the Interconversion of Testosterone to Androstenedione (4-dione)
Description:  A radioactive assay was used for the enzyme kinetics in the presence of EM1404 at different concentrations for its Ki determination. Reactions were initiated by the addition of enzyme to the reaction mixture containing substrate and test compounds. After the reaction, the steroids were extracted, separated by TLC migration, and analyzed by phosphorimaging. The Km for testosterone oxidation in the absence of EM-1404, and four apparent Km values in the presence of 1, 3, 10, and 25 nM EM-1404, were determined. Ki value was further determined by a plot of Km vs. the inhibitor concentration.
Affinity data for this assay
 

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Last update November 1, 2007
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