Assay Method Information

Assay Name:  AChE and BChE Inhibition Activity
Description:  Inhibitory activities of AChE and BChE were evaluated by using the Ellman's method [34]. Herein compounds 2(a-h) and 5(a-h) were evaluated as inhibitors of AChE and BChE. For initial screening each compound was dissolved in DMSO (end concentration of DMSO was less than 1% in assay) and tested at a final concentration of 0.5 mM. The compounds with considerable inhibition (more than 50%) were subjected to further analysis by making their 8 to 10 serial dilutions in assay buffer (50 mM Tris-HCl, 0.1 M NaCl and 0.02 M MgCl2, pH 8.0). Reaction mixture comprised of 20 µL assay buffer, 10 µL of test compound, 10 µL of 0.031 U/mL of enzyme (0.5 and 3.4 U/mg of AChE or BuChE respectively). This mixture was incubated at 25 °C for 10 min. After this preincubation, enzymatic reaction was started by addition of 10 µL of 1 mM acetylethiocholine iodide or butyrylthiocholine chloride (depending on the enzyme) and the mixture was incubated again for 15 min. The amount of enzymatic product was esti
Affinity data for this assay
 

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