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Assay Method Information

Assay Name:  Cholinesterase Activity Assay
Description:  Reactions were performed in a mediumcontaining substrate (0.05-0.4 mM) combined with 0.125 mM DTNB in 100 mM 3-(N-morpholino)propanesulfonic acid buffer, pH 8.0 at 25 °C, initiated by the addition of enzyme, and monitored spectrophotometrically at 412 nm in a UV-visible 1700 Shimadzu PC spectrophotometer. The activity was determined by measuring the increase in absorbance at 412 nm for 7 min (εTNB = 14.2 mM^-1 cm^-1). Activities of the treated samples were compared to control to estimate percentage of inhibition (I%). Dose-response curves were plotted using the GraphPad Prism 5 software and IC50 values were determined. Data are expressed as mean ± SEM. An enzyme kinetic assay was performed for compound 2 at different butyrylthiocholine (BTC) concentrations (0.05-0.3 mM). Donepezil-HCl (Sigma) was tested as a reference compound.
Affinity data for this assay
 

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Last update November 1, 2007
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