Table 2.
Summary of experimental conditions for determination of affinities at recombinant human monoamine receptors in vitro
| Receptor | Cell line | [3H]Radioligand (nM) | Kd (nM) | Non-specific (μM) | Inc. buffer | Inc. time (min) & temp. (°C) | Literature reference |
|---|---|---|---|---|---|---|---|
| hD2S | CHO | Spiperone (0.6) | 0.084 | (+)butaclamol (1) | A | 120, 37° | Cussac et al. (2000) |
| hD2L | CHO | Spiperone (0.6) | 0.035 | (+)butaclamol (1) | A | 120, 37° | Cussac et al. (2000) |
| hD3 | CHO | Spiperone (0.4) | 0.21 | Raclopride (10) | B | 160, 25° | Cussac et al. (2000) |
| hD4.4 | CHO | Spiperone (0.6) | 0.15 | Haloperidol (1) | A | 120, 37° | Newman-Tancredi et al. (1997) |
| hD1 | CHO | SCH23390 (0.1) | 0.38 | SKF38393 (10) | C | 120, 25° | Pedersen et al. (1994) |
| h5-HT1A | HeLa | 8-OH-DPAT (1) | 0.71 | 5-HT (10) | D | 30, 25° | Newman-Tancredi et al. (2005) |
| h5-HT1B | COS7 | 5-CT (3) | 0.65 | 5-HT (10) | D | 30, 25° | Pauwels et al. (1997) |
| h5-HT1D | COS7 | 5-CT (3) | 0.60 | 5-HT (10) | D | 30, 25° | Pauwels et al. (1997) |
| h5-HT2A | CHO | Ketanserin (0.5) | 0.24 | 5-HT (10) | A | 120, 23° | Millan et al. (2002a) |
| h5-HT2B | CHO | Mesulergine (2) | 1.68 | 5-HT (10) | A | 120, 23° | Millan et al. (2002a) |
| h5-HT2C | CHO | Mesulergine (2) | 0.56 | 5-HT (10) | A | 120, 23° | Millan et al. (2002a) |
| h5-HT7A | HEK293 | 5-CT (1.5) | 1.66 | 5-HT (10) | D | 90, 37° | Bard et al. (1993) |
| hα2A | C6 glial | RX821002 (2) | 1.16 | phentolamine (10) | E | 60, 25° | Wurch et al. (1999) |
| hα2B | C6 glial | RX821002 (10) | 9.27 | phentolamine (10) | E | 60, 25° | Wurch et al. (1999) |
| hα2C | C6 glial | RX821002 (4) | 2.21 | phentolamine (10) | E | 60, 25° | Wurch et al. (1999) |
Abbreviations: CHO, Chinese hamster ovary cells; HEK293, human embryonic kidney cells 293.
Buffer A: HEPES 20 mM (pH 7.4), NaCl 120 mM, KCl 5 mM, EDTA 1 mM, MgCl2 5 mM; Buffer B: Tris-HCl 50 mM (pH 7.6), NaCl 120 mM, KCl 5 mM; CaCl2 2 mM, MgCl2 5 mM, BSA 0.1%; Buffer C: HEPES 20 mM (pH 7.2); Buffer D: Tris-HCl 50 mM (pH 7.6), CaCl2 4 mM, pargyline 10 μM, ascorbic acid 0.1%; Buffer E: Tris-HCl 50 mM (pH 7.6).