| Assay Method Information | |
| | AChE/BChE Inhibitory Assay |
| Description: | AChE and BChE inhibitory assay was carried out by following Ellman's methodology [Ellman et al., Biochem. Pharmacol., 7:88-95] using AChE (Electric eel type-VI-S, Sigma-Aldrich GmbH USA, code 1001596210), BChE (Equine serum Lyophilized Sigma-Aldrich GmbH USA, code 101292670) and DTNB (Sigma-Aldrich Germany, code 101261619), which produced colored product (5-thio-2-nitrobenzoate) whose concentration can be measured by the increase in absorbance at 412 nm using mQuant microplate spectrophotometer (MQX200, BioTek USA). Other reagents, like Acetylthiocholine iodide (Sigma-Aldrich UK, code 101303874), Butyrylthiocholine Iodide (Sigma-Aldrich Switzerland, code 101334643) were employed. Galantamine hydrobromide Lycoris Sp. (Sigma-Aldrich France, code G1660) and Donepezil were used as reference drugs. Stock solution of the synthesized quinazoline was prepared with 0.1 M phosphate buffer (KH2PO4/K2HPO4) having of pH 8.0. Appropriate amount of DTNB (Ellman's reagent), quinazoline compounds, 0.03 U/ml of enzymes (AChE and BChE) were reacted by pre-incubating at 30 °C for 10 min and then further incubating for 15 min after addition of 1 mM ATCI or BTCI. |
| Affinity data for this assay | |
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