| Assay Method Information | |
| | AChE and BChE Inhibition Assay |
| Description: | In vitro anti-AChE activity was performed based on the modified Ellman's method [Ellman et al., Biochem. Pharmacol., 71:88-95]. For this purpose, synthesized compounds 6 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 μL potassium phosphate buffer (KH2PO4/K2HPO4, 0.1 M, pH 8), 25 μL prepared sample as described above, 25 μL enzyme with final concentration of 0.22 U/mL in buffer. They were preincubated for 15 min at room temperature, and then 125 L DTNB (3 mM in buffer) was added. Characterization of the hydrolysis ofATCI catalyzed by AChE 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). A control experiment was performed under the same conditions without inhibitor and the blank contained buffer, water, DTNB, and substrate. The described method was also used for BChE inhibition assay. For all compounds, four different concentrations were tested for each compound in triplicate to obtain the range of 20%−80% inhibition for AChE. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |