| Assay Method Information | |
| | AChE and BuChE Inhibition Assay |
| Description: | The reaction mixture was consisted of 100 µL of the total volume. To the solution of PBS (pH 8.0, 30 µL) consisted of KH2PO4 and K2HPO4 in 96-well plate, the solution of enzyme AChE (15 µL, 0.132 U/mL or BuChE (15 µL, 0.04 U/mL) in the PBS was added and followed by the addition of the synthesized compounds (20 µmol/L, 25 µL DMSO). The resulting mixture were thoroughly mixed, pre-incubated for 10 min at 37 °C and pre-read at 412 nm. The reaction started by the addition of 15 µL of 4mM well-1substrate (acetylthiocholine iodide-ATCI or butyrylthiocholine chloride-BuSCH), followed by the incorporation of 15 µL of 6 mM well-1 Ellman solution (DTNB). After 30 min of incubation at 37 °C, absorbance was recorded at 412 nm by 96-well plate reader Bio-Rad Laboratories Inc. All experiments were performed with their respective standards (controls) in triplicate. As a positivecontrol Huperzine A (0.8 µM well-1) or Iso-OMPA (16.0 µM well-1) was practiced. |
| Affinity data for this assay | |
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