| Assay Method Information | |
| | In Vitro Functional Assay of Muscarinic Acetylcholine Receptor Activity |
| Description: | Primary compound plates were prepared in 100% DMSO in opaque 96-well plates (VWR) and serially diluted in half log increments. Secondary compound plates were prepared at 3× concentration in assay buffer. Compounds were added to white, clear bottom tissue culture treated 96-well plates (Fisher Scientific). Coelenterazine-loaded cells were then added at 5×105 cells/well.Compounds and cells were incubated at room temperature for 30 min in the dark. Acetylcholine at the EC80 concentration was added and calcium flux measured using a FlexStation 3 (Molecular Devices). Sigmoidal dose-response curves were generated by measuring luminescence over 40 sec and calculating the area under the curve. Dose response curves and |
| Affinity data for this assay | |
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