Specific conditions used for studies of displacement of radioligands by the compounds under study
| Assay | Radiolabel | Tissue | Incubation Buffer | Incubation | Nonspecific Binding |
|---|---|---|---|---|---|
| σ1R | 3.0 nM [3H](+)-pentazocine (Perkin-Elmer, Boston, MA) | 8.0 mg/tube, frozen guinea pig brains excluding the cerebella (Pel Freez Biologicals, Rogers, AR) | 10 mM Tris-HCl with 0.32 M sucrose, pH 7.4 | 120 min at room temperature | 10 μM haloperidol |
| σ2R | 3.0 nM [3H]DTG (Perkin-Elmer) with 200 nM (+)-pentazocine | 8.0 mg/tube, frozen guinea pig brains excluding the cerebella (Pel Freez Biologicals) | 10 mM Tris-HCl with 0.32 M sucrose, pH 7.4 | 120 min at room temperature | 100 μM haloperidol |
| DAT | 0.50 nM [3H]WIN 35,428 (Perkin-Elmer) | 1.0 mg/tube, frozen striatum (from male Sprague-Dawley rats brains supplied on ice; Bioreclamation, Hicksville, NY) | Modified sucrose phosphate buffer (0.320 M sucrose, 7.74 mM Na2HPO4, 2.26 mM NaH2PO4, pH adjusted to 7.4) | 120 min on ice | 100 µM cocaine HCl |
WIN 35,428, (−)-2-β-carbomethoxy-3-β-(4-fluorophenyl)tropane-1,5-napthalenedisulfonate.