Assay Method Information

Assay Name:  Scintillation Proximity Assay
Description:  [125I]-()DOI binding experiments are carried out in SPA 96-well format. Membranes used in this assay are prepared from AV-12 cells stably expressing recombinant 5-HT2C receptor (human). The incubation is initiated by the addition of a mixture of WGA PVT SPA beads (0.5 mg/well, Perkin Elmer (MA, USA), RPNQ0001) and 2.5 μg membranes to assay buffer (50 mM Tris-HCl, 10 mM MgCl2, 0.5 mM EDTA, 10 μM pargyline, 0.1% ascorbic acid, pH7.4) containing 0.2 nM [[125I]-()DOI and varying concentrations of the test compound (10 point concentration response curves). Non-specific binding is determined in the presence of 20 μM 1-(1-Naphthyl) piperazine. Samples are incubated for 4 hr. at room temperature (22 C.) and then read in a Microbeta Trilux.
Affinity data for this assay
 

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