The experimental conditions used in the receptor binding assays
| Receptor | Ligand (Kd, nM) | Tissue (conc mg w.w./ml incubation volume) or cell (conc mg protein/ml incubation volume) | Incubation Time and Temperature (min/°C) | Incubation Buffer (in mM), (pH) | Nonspecific Binding (μM) |
|---|---|---|---|---|---|
| 5-HT1A | [3H]8-OH-DPAT (0.65) | Rat hippocampus (2/2) | 45 /37* | 50 Tris-HCl, 2 CaCl, 1 MgCl2, 1 MnCl2 (pH 7.4) | 10 WAY100635 |
| 5-HT1B | [3H]Cyanopindolol (0.05) | Rat cortex (1.5/0.25) | 90 /37 | 10 Tris-HCl, 154 NaCl, 0.01 pargyline, 0.06 (±)isoprenaline (pH ?) | 10 5-HT |
| 5-HT2A | [3H]Ketanserin (0.60) | Rat cortex (2.5/2) | 45 /37 | 50 Tris-HCl (pH 7.7) | 10 methysergide |
| 5-HT6 | [3H]5-HT (3.00) | CHOr5-HT6 (0.2/2) | 30 /30 | 50 Tris-HCl, 4 MgCl2, 1 EDTA (pH 7.4) | 10 methiothepin |
| 5-HT7 | [3H]5-HT (0.40) | CHOr5-HT7 (0.04/1) | 60 /30 | 50 Tris-HCl, 4 MgCl2, 1 EDTA (pH 7.4) | 10 methiothepin |
| 5-HT uptake | [3H]Citalopram (0.78) | Rat cortex (4/2) | 120 /25 | 50 Tris-HCl, 5 KCl, 120 NaCl (pH 7.4) | 0.5 paroxetine |
| α1 | [3H]Prazosin (0.04) | Rat cortex (2.5/2) | 60 /25 | 50 Tris-HCl, 0.1% a.a.1-b (pH 7.7) | 100 phentolamine |
| α2 | [3H]RX821002 (0.40) | Rat cortex (5/2) | 60 /25 | 50 Tris-HCl, 0.1% a.a. (pH 7.7) | 100 phentolamine |
| β | [3H]DHA (0.50) | Rat cortex (5/2) | 60 /25 | 50 Tris-HCl, 120 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 0.01 pargyline, 0.1% a.a. (pH 7.6) | 10 (±)alprenolol |
| Muscarinic | [3H]QNB (0.17) | Rat cortex (0.5/2) | 60 /25 | 50 Tris-HCl, 120 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 0.01 pargyline, 0.1% a.a. (pH 7.6) | 100 oxotremorine |
| Nicotinic | [3H]Nicotine (0.34) | Rat brain-cerebellum (3.2/0.2) | 10 /25 | 20 HEPES, 118 NaCl, 4.8 KCl, 2.5 CaCl2, 1.2 MgSO4 (pH 7.4) | 10 nicotine |
| D1 | [3H]SCH23390 (0.29) | Rat striatum (2.5/2) | 60 /25* | 50 Tris-HCl, 120 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 0.01 pargyline, 0.1% a.a. (pH 7.6) | 1 flupentixol |
| D2 | [3H]Raclopride (1.20) | LtkhD2A (0.03/0.5) | 60 /25 | 50 Tris-HCl, 120 NaCl, 5 KCl, 1.5 CaCl2, 4 MgCl2, 1 EDTA (pH 7.4) | 1 butaclamol |
| D3 | [3H]Raclopride (1.60) | CHOhD3 (0.03/0.5) | 60 /25 | 50 Tris-HCl, 120 NaCl, 5 KCl, 1.5 CaCl2, 4 MgCl2, 1 EDTA (pH 7.4) | 1 butaclamol |
| Histamine H1 | [3H]Pyrilamine (0.73) | Rat cortex (5.3/0.2) | 25 /25 | 50 Na2HPO4/KH2PO4 (pH 7.4) | 100 pyrilamine |
| Histamine H2 | [3H]Tiotidine (17) | Guinea pig cortex (5.3/0.2) | 30 /25 | 50 Na2HPO4/KH2PO4 (pH 7.4) | 100 cimetidine |
| GABAA | [35S]TBPS (70) | Rat cortex (3.2/0.2) | 90 /25 | 50 Tris-citrate, 200 NaCl (pH 7.4) | 4.75 TBPS |
| NMDA | [3H]MK801 (11.99) | Rat cortex | 15 /25 | 5 Tris-HCl, 0.01 glutamate, 0.01 glycine, 0.1 Spermidine (pH 7.4) | 100 MK801 |
| AMPA | [3H]AMPA (82) | Rat forebrain (8/0.2) | 60 /4 | 50 Tris-HCl (pH 7.4) | 1000 glutamate |
| Benzodiazepine | [3H]Flunitrazepam (2.1) | Rat cortex (2.7/0.2) | 30 /0 | 50 Tris-HCl (pH 7.4) | 100 diazepam |
| Galanin | [125I]Galanin (0.15) | Rat hypothalamus (0.75/0.2) | 40 /37 | 20 HEPES, 2.5 MgCl2, 0.5 EDTA, 0.05% bovine serum albumin, 1 mg/ml bacitracin (pH 7.4) | 1 galanin |
* Preincubated 10 min at 37°C.
↵1-b a.a., ascorbic acid.