| Assay Method Information | |
| | Radioligand Binding Assay |
| Description: | Affinity at transmembrane glutamate metabotropic mGluR5 receptor subtypes was evaluated according to the methods of Anderson (Anderson et al., J Pharmacol. Exp. Ther., (2002), Vol. 303(3), pp. 1044-51), with some modifications. Cloned mGluR5 was obtained by re-suspending CHO T-REx h-mGluR5 cells (50 μg/well) in 20 mM HEPES, 2 mM MgCl2, 2 mM CaCl2, pH 7.4, that then were incubated in a final volume of 1 ml for 60 min at 25° C. with 4 nM [3H]MPEP in the absence or presence of competing drugs. Non-specific binding was determined in the presence of 10 mM MPEP. The incubation was stopped by addition of cold Tris buffer pH 7.4 and rapid filtration through 0.2% polyethyleneimine pretreated Filtermat 1204-401 (Perkin Elmer) filters. The filters were then washed with cold buffer and the radioactivity retained on the filters was counted by liquid scintillation spectrometry (Betaplate 1204 BS-Wallac). |
| Affinity data for this assay | |
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