Solution Edos and you can Post was indeed measured playing with Estradiol EIA Kit (Cayman Chemicals, MI) and you may androstenedione chemical immunoassay package (Oxford Biomedical Look Inc., MI), correspondingly. ELISAs out-of T and E1 was did depending on the same means of 17,20?-P (Matsuyama mais aussi al., 1998b) and you will explained lower than temporarily.

Stock solutions: Coating buffer, 0.05 M carbonate buffer pH 8.4, containing 0.05% NaN3; washing solution, 0.85% NaCl; blocking solution, 0.05 M PBS containing 0.1% BSA, 3% sucrose, and 0.005% thimerosal; assay buffer, 0.05 M borate buffer, pH 7.8, containing 0.1% BSA and 0.01% thimerosal; substrate solution, 0.2 M citrate buffer, pH 4.5, containing 0.01% H2O2, added 0.5% o-phenylenediamine immediately before use; stopping solution, 6 N H2SO4. All the solutions except for the stopping solution were stored at 4°C.

2nd Antibody-coated Microtiter Plates: Microtiter dishes (MS-3596 F/H plate, Sumitomo Bakelite Co.) was basically decorated with 100 ?l each well out of goat anti-bunny IgG (fifteen ?g/ml inside the coating buffer). Continue reading