| Assay Method Information | |
| | 5α-Reductase Activity Assay |
| Description: | The reaction mixtures contained a final volume of 1 mL, 1 mM DTT, sodium phosphate buffer 40 mM, at pH 6.5, 2 mM, NADPH, 2 nM [1,2,6,7-3H]T or [1,2,3,7-3H(N)] 4-dione and the prostatic enzyme fractions. The amount of prostatic enzyme fractions was determined to adjust the rate of conversion of T to DHT or 4-dione to 5α-dione to around 28%. T, DHT, unlabeled 4-dione or 5α-dione concentrationswere adjusted to 25-250 nM in the incubating medium, by adding one of the following reagents cold T, DHT, unlabeled 4-dione or 5α-dione. The reactions in duplicate were started when added to the enzymatic fraction (400 μg protein in a volume of 80 μL) incubated at 37°C for 60 min13 and stopped by mixing with 1 mL ofdichloromethane. Incubation without tissue was used as a control. All reactions were carried out in two different times by duplicate. |
| Affinity data for this assay | |
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