| Assay Method Information | |
| | Radioligand Binding Assay (IC50) and Functional Assay (EC50/IC50) |
| Description: | Compounds were evaluated for their binding affinity to the membranes of CHO-K1 cells expressing human MCH1R (hMCH1R) in a competition binding assay with [125I]-MCH as the radioligand. After incubation, the membranes were filtrated onto GF/C filter plates and dried. The radioactivity was counted using a microplate scintillation counter (TopCount, PerkinElmer). Non-specific binding was determined by including 1 uM unlabeled MCH in the binding reaction. Antagonistic activity (EC50/IC50) was estimated by the inhibitory effect of compounds on intracellular calcium mobilization induced by MCH using a FLIPR in CHO-K1 cells expressing human MCH1R. The cells were treated with the test compounds for 5 min before addition of MCH. |
| Affinity data for this assay | |
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