Competition binding conditions at recombinant, human serotonin (5-HT) receptors, at recombinant, human hM1 and at native histamine H1 receptors
| Receptor | Tissue | Radioligand (nM) | Buffer ingredients (mM) | pH | Time, temperature | Non-specific (μM) | Kd |
|---|---|---|---|---|---|---|---|
| nM | |||||||
| h5-HT1A | CHO cells | [3H]8-OH-DPAT (0.4) | HEPES (20), MgSO4(5) | 7.5 | 150 min, 22°C | 5-HT (10) | 0.65 |
| h5-HT1B | CHO cells | [3H]GR125,743 (1.0) | Tris (50), CaCl2 (4), Asc.Ac. (0.1% w/v) | 7.7 | 60 min, 25°C | 5-HT (10) | 1.27 |
| h5-HT1D | CHO cells | [3H]GR125,743 (1.0) | Tris (50), CaCl2 (4), Asc.Ac. (0.1% w/v) | 7.7 | 60 min, 25°C | 5-HT (10) | 0.67 |
| h5-HT2A | CHO cells | [3H]Ketanserin (0.5) | HEPES (20), EDTA (2), Asc.Ac. (0.1% w/v) | 7.7 | 40 min, 22°C | Mianserin (1) | 0.22 |
| h5-HT2B | CHO cells | [3H]Mesulergine (1.0) | HEPES (20), EDTA (2), Asc.Ac. (0.1% w/v) | 7.7 | 60 min, 22°C | 5-HT (10) | 1.44 |
| h5-HT2C | CHO cells | [3H]Mesulergine (1.0) | HEPES (20), EDTA (2), Asc.Ac. (0.1% w/v) | 7.7 | 120 min, 22°C | Mianserin (1) | 0.55 |
| H1 | Guinea pig cerebellum | [3H]Pyrilamine (0.5) | Na2HPO4/KH2PO4(50) | 7.4 | 10 min, 25°C | Triprolidine (100) | 0.37 |
Asc.Ac., ascorbic acid.