| Assay Method Information | |
| | [35S]TBPS Binding Assay |
| Description: | Briefly, aliquots of membrane solution (0.5 mg/mL final protein concentration in assay) were incubated with 5 μM GABA, 2 nM [35S]TBPS (45−120 Ci/mmol), and 5 μL aliquots of steroid in DMSO solution (final assay concentrations ranged from 1 nM to 10 μM), and brought to a final volume of 1 mL with 200 mM NaCl, 50 mM potassium phosphate buffer, pH 7.4. Control binding was defined as binding observed in the presence of 0.5% DMSO and the absence of steroid. Nonspecific binding was defined as binding observed in the presence of 200 μM picrotoxinin and ranged from 6.1 to 14.3% of total binding. Assay tubes were incubated for 2 h at room temperature. |
| Affinity data for this assay | |
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