title: Molybdate stabilizes soluble [3H]dexamethasone binding sites in rat brain tissue in incubations at room temperature (22 degrees C).
abstract: We have described an in vitro procedure for the measurement of glucocorticoid receptor concentrations in soluble fractions prepared from rat brain tissue using a relatively short incubation period of 1 h at room temperature (22 degrees C). Using [3H]dexamethasone as ligand we found that, with the addition of sodium molybdate (Na2MoO4) to the buffer, estimates of receptor-binding parameters derived from Scatchard analysis (Kd and Bmax) did not differ significantly between assays using 4-5 h incubations at 0-4 degrees C and those using a 1 h incubation performed at room temperature (22 degrees C). The use of molybdate was critical; 1 h incubations at 22 degrees C in the absence of molybdate resulted in a substantial (approximately 65%) loss of [3H]dexamethasone binding.
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D003907:Dexamethasone / Q000378:metabolism*; D005260:Female; D008297:Male; D008982:Molybdenum / Q000494:pharmacology*; D011869:Radioligand Assay; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000032:analysis; D013696:Temperature
chemical_list: D011965:Receptors, Glucocorticoid; D003907:Dexamethasone; D008982:Molybdenum; C024687:sodium molybdate(VI)