Table 1.

Radioligand Binding Procedure

Radioligand	Competing Ligand	Pre-Incubation	Incubation	Wash
[3H]Pirenzepine 10 nM	1 μM Atropine	Krebs-Hepes pH 7.4 10 min 22°C	Krebs-Hepes pH 7.4 60 min 22°C	Krebs-Hepes pH 7.4 2 × 2 min 1 dip dH2O 4°C
[3H]AF-DX 384 7 nM	1 μM Atropine	Krebs-Hepes pH 7.4 10 min 22°C	Krebs-Hepes pH 7.4 120 min 22°C	Krebs-Hepes pH 7.4 2 × 2 min 1 dip dH2O 4°C

^*Krebs-Hepes: 0.1 M NaCI, 2.0 mM KCI, 1.2 mM KH2PO4, 1.2 mM MgC12, 2.5 mM CaCI2, 5.0 mM HEPES, 5.5 mM glucose