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Assay Method Information

Assay Name:  Cholinesterase Inhibition Assay
Description:  Suitable agents like donepezil hydrochloride and tacrine hydrochloride hydrate were used as standards for this study. Buffer solution (50 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 0.02 M MgCl26H2O) was used to dilute the stock solutions of the test compounds dissolved in minimum volume of DMSO (1%). First of all 50 L of AChE (0.22 U/mL prepared in 50 mM Tris-HCl, pH 8.0, 0.1% w/v bovine serum albumin, BSA) or 50 L of BuChE (0.06 U/mL prepared in 50 mM Tris-HCl, pH 8.0, 0.1% w/v BSA) and 10 L of different concentrations of the test and standard compounds (0.001-100 M) were incubated in 96-well plates at room temperature for 30 min. Further, 30 L of the substrate viz. Acetylthiocholine iodide [ATCI (15 mM)] or butyrlthiocholine iodide [BTCI (15 mM)] was added and incubated for 30 min. Finally 160 L DTNB (1.5 mM) was added and absorbance was measured at 415 nm wavelength.
Affinity data for this assay
 

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Last update November 1, 2007
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