Testis-Like Steroidogenesis: MATERIALS AND METHODS(3)

19 Apr
2013

MATERIALS AND METHODS(3)

RIA

The concentration of testosterone in plasma and gonadal homogenates was determined by RIA as described in detail by Licht et al.. Briefly, plasma samples and gonadal homogenates were extracted twice with diethyl ether (Mal-linkrodt, St. Louis, MO), and testosterone was measured with an antiserum cospecific for testosterone and dihydrotestosterone. [1,2,6,7-3H]Testosterone was used as the label (NEN). Measurements of plasma androstenedione were determined by RIA as described by Glickman et al.. Plasma samples were extracted as above and assayed with an antiserum specific for androstenedione with [1,2,6,7-3H]androstenedione as label (NEN). birth control yasmin

Protein Assay

Protein content for each sample was determined by Bradford protein assay: 160 ^l of serial dilutions of the gonadal homogenates was added to a microtiter tray with 40 ^l of protein assay dye reagent (Bio-Rad, Hercules, CA) according to the manufacturer’s instructions. Serial dilutions of BSA were used as the standards. Absorbance was determined by a Bio-Rad UV microplate reader.

Statistics

Significance between groups was assessed with the non-parametric Mann-Whitney U Test and are presented as means ± SD.

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