| Assay Method Information | |
| | AChE and BChE Inhibition Assay |
| Description: | In vitro AChEI activity was performed based on the modified Ellman's methodas previously reported by our group using a 96-well plate reader (BioTek ELx808). For this purpose, compounds 8 were dissolve in a mixture of DMSO (5 mL) and methanol (5 mL) and diluted in 0.1 M KH2PO4/K2HPO4 buffer (pH 8.0). Each well contained 50 uL potassium phosphate buffer (KH2PO4/K2HPO4, 0.1 M, pH 8), 25 uL prepared sample as described above, 25 uL enzyme with final concentration of 0.22 U/mL in buffer. They were preincubated for 15 min at rt, and then 125 uL DTNB (3 mM in buffer) was added. Characterization of the hydrolysis of ATCI catalyzed byAChE was performed spectrometrically at 405 nm followed by the addition of substrate (ATCI 3 mM in water). The change in absorbance was measured at 405 nm after 15 min. The IC50 values were determined graphically from inhibition curves (log inhibitor concentration vs. percent of inhibition). |
| Affinity data for this assay | |
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