Incubation conditions and ligand specifications for receptor autoradiography
| Receptor . | Alpha-1 . | Alpha-2 . |
|---|---|---|
| [3H]-Ligand | [3H]-Prazosin | [3H]-UK-14,304 |
| Pharmacology | Antagonist | Agonist |
| Ligand concentration (L) | 0.2 nM | 0.64 nM |
| KD value | 0.2 nM | 1.4 nM |
| Specific activity SA | 80.5 Ci/mmol | 81.2 Ci/mmol |
| Displacer | Phentolamine mesylate | Phentolamine mesylate |
| Displacer concentration | 10 μM | 10 μM |
| Incubation buffer | 50 mM Na/K-phosphate buffer (pH 7.4) | 50 mM Tris-HCl + 100 μM MnCl2 (pH 7.7) |
| Preincubation | 15 min, 22°C | 15 min, 22°C |
| Main incubation | 60 min, 22°C | 90 min, 22°C |
| Rinsing | 1) 2 × 5 min, 4°C 2) distilled water, 1 × 22°C | 1) 5 min, 4°C 2) distilled water, 1 × 22°C |
| Receptor . | Alpha-1 . | Alpha-2 . |
|---|---|---|
| [3H]-Ligand | [3H]-Prazosin | [3H]-UK-14,304 |
| Pharmacology | Antagonist | Agonist |
| Ligand concentration (L) | 0.2 nM | 0.64 nM |
| KD value | 0.2 nM | 1.4 nM |
| Specific activity SA | 80.5 Ci/mmol | 81.2 Ci/mmol |
| Displacer | Phentolamine mesylate | Phentolamine mesylate |
| Displacer concentration | 10 μM | 10 μM |
| Incubation buffer | 50 mM Na/K-phosphate buffer (pH 7.4) | 50 mM Tris-HCl + 100 μM MnCl2 (pH 7.7) |
| Preincubation | 15 min, 22°C | 15 min, 22°C |
| Main incubation | 60 min, 22°C | 90 min, 22°C |
| Rinsing | 1) 2 × 5 min, 4°C 2) distilled water, 1 × 22°C | 1) 5 min, 4°C 2) distilled water, 1 × 22°C |
Incubation conditions and ligand specifications for receptor autoradiography
| Receptor . | Alpha-1 . | Alpha-2 . |
|---|---|---|
| [3H]-Ligand | [3H]-Prazosin | [3H]-UK-14,304 |
| Pharmacology | Antagonist | Agonist |
| Ligand concentration (L) | 0.2 nM | 0.64 nM |
| KD value | 0.2 nM | 1.4 nM |
| Specific activity SA | 80.5 Ci/mmol | 81.2 Ci/mmol |
| Displacer | Phentolamine mesylate | Phentolamine mesylate |
| Displacer concentration | 10 μM | 10 μM |
| Incubation buffer | 50 mM Na/K-phosphate buffer (pH 7.4) | 50 mM Tris-HCl + 100 μM MnCl2 (pH 7.7) |
| Preincubation | 15 min, 22°C | 15 min, 22°C |
| Main incubation | 60 min, 22°C | 90 min, 22°C |
| Rinsing | 1) 2 × 5 min, 4°C 2) distilled water, 1 × 22°C | 1) 5 min, 4°C 2) distilled water, 1 × 22°C |
| Receptor . | Alpha-1 . | Alpha-2 . |
|---|---|---|
| [3H]-Ligand | [3H]-Prazosin | [3H]-UK-14,304 |
| Pharmacology | Antagonist | Agonist |
| Ligand concentration (L) | 0.2 nM | 0.64 nM |
| KD value | 0.2 nM | 1.4 nM |
| Specific activity SA | 80.5 Ci/mmol | 81.2 Ci/mmol |
| Displacer | Phentolamine mesylate | Phentolamine mesylate |
| Displacer concentration | 10 μM | 10 μM |
| Incubation buffer | 50 mM Na/K-phosphate buffer (pH 7.4) | 50 mM Tris-HCl + 100 μM MnCl2 (pH 7.7) |
| Preincubation | 15 min, 22°C | 15 min, 22°C |
| Main incubation | 60 min, 22°C | 90 min, 22°C |
| Rinsing | 1) 2 × 5 min, 4°C 2) distilled water, 1 × 22°C | 1) 5 min, 4°C 2) distilled water, 1 × 22°C |
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