Nuclear Androgen Receptors in the Mouse Kidney: Validation of a New Assay*

摘要: A method for determining nuclear androgen receptors in mouse kidney was developed using [3H]methyltrienolone as the labeled ligand. Pyridoxal 5′-phosphate used to extract from nuclei isolated with hexylene glycol technique. The pyridoxal extraction yielded receptor concentrations about twice high those measured after 0.4 M KCl or by a direct exchange reaction. recovery of also increased 4-fold use initial homogenization medium rather than Tris-buffer containing glycerol, which is cytosol assays. complete endogenously bound testosterone achieved during an 18-h incubation at 4 C. had KD 5 nM [3H] methyltrienolone, specific (5α-dihydrotestosterone = methyltrienolone > estradiol rogesterone » dexamethasone aldosterone) and sedimented 4S enti...