| Assay Method Information | |
| | Anticholinesterase Enzyme Inhibition Assay |
| Description: | The in vitro AChE inhibitory activity was measured using the methods described earlier [Biochem. Pharmacol. 7:88-95]. Briefly, stock solutions (1mg/mL) of test compounds were prepared using DMSO. Working solutions (0.01-100痢/mL) were prepared by serial dilutions. The various concentrations of test compounds (10 無) were pre-incubated with sodium phosphate buffer (0.1M; pH 8.0; 150 無), and AChE solution (0.1 U/mL; 20 無) for 15 min at 25 C. The reaction was initiated by addition of DTNB (10 mM; 10 無) and ATChI (14 mM; 10 無). The reaction mixture was mixed using a cyclomixer and incubated for 10min at room temperature. The absorbance was measured using a microplate reader at 410 nm wavelength against the blank reading containing 10無 DMSO instead of test compound. The % inhibition was calculated using the formula described in Eq. (1) and the IC50 was calculated. Donepezil (0.01-100痢/mL) was used as the positive control. |
| Affinity data for this assay | |
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