| Assay Method Information | |
| | AChE and BChE Inhibition Assay |
| Description: | A 96-microliter well plate was used for screening purpose. Each pyridopyrazine derivative was first dissolved in dimethyl sulfoxide to prepare the stock solutions. The synthetic compounds were then screened against electric eel acetylcholinesterase (eeAChE) and equine butyrylcholinesterase (BChE) at different concentrations (1 µM or 10 µM) of dimethyl sulfoxide as negative control in primarily screening. The compounds which showed ≥ 50% inhibition were further evaluated by making their serial dilutions (~ 6 to 7 times) with buffer (50 mM Tris-HCl, 0.02 M MgCl2 6H2O, and 0.1 M NaCl at pH 8.0). The reaction mixture which comprised of buffer solution (20 µL), synthetic compound (10 µL) and enzyme (0.5 and 3.4 U/mg, eeAChE or equine BChE) was primarily incubated for ten minutes at room temperature (25 C). Subsequently to the mixture, once the preincubation over, the acetylthiocholine iodide or butyrylthiocholine chloride (10 μL of 10 mM) was added and incubated it further for |
| Affinity data for this assay | |
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