| Assay Method Information | |
| | M3 Receptor Affinity Test |
| Description: | The experiment was performed by a radioisotope affinity assay. The M3 receptor membrane protein was prepared at 10 μg/mL by Biology Department of WuXi AppTec, and compounds were serially diluted in a 2.5-fold gradient in DMSO to obtain 10 concentrations. Radioisotope 3H-NMS was prepared at 0.2 nM in an assay buffer (10 mM HEPES, 1 mM MgCl2, pH 7.40). In formal testing, the experimental system comprising 1 of the test compound, 100 μL of M3 receptor membrane protein and 100 μL of radioisotope was shaken in a shaker at 300 RPM for 2 h at room temperature. A GF/C plate (Perkin Elmer, Cat No. 6055690) was prefoamed with 0.3% PEI, and the membrane proteins in the reaction solution were collected by filtration onto the GF/C plate. The signal values were read by a Perkin Elmer Microbeta2 instrument and the inhibition rate for each concentration point was shown as a percentage. |
| Affinity data for this assay | |
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