| Assay Method Information | |
| | Acetylcholinesterase Assay |
| Description: | The AChE inhibition activity was determined according to the Ellman's method with slight modifications. Total volume of the reaction mixture was 100 µL. It contained 60 µL Na2HPO4 buffer with concentration of 50 mM and pH 7.7. Ten µL test compound (0.5 mM well-1) was added, followed by the addition of 10 µL (0.005 unit well-1) enzyme. The contents were mixed and preread at 405 nm. Then contents were pre-incubated for 10 min at 37 °C. The reaction was initiated by the addition of 10 µL of 0.5 mM well-1 substrate (acetylthiocholine iodide), followed by the addition of 10 µL DTNB (0.5 mM well-1). After 30 min of incubation at 37 °C, absorbance was measured at 405 nm. Synergy HT (BioTek, USA) 96-well plate reader was used in all experiments. All experiments were carried out with their respective controls in triplicate. |
| Affinity data for this assay | |
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