| Assay Method Information | |
| | AChE and BuChE Inhibition Assay |
| Description: | This spectrophotometric assay is based on the reaction of 5,5-dithio-bis(2-nitrobenzoic)acid (DTNB) with thiocholine to yield a colored product. Shimadzu (Kyoto, Japan) UV-1700 UV-vis spectrophotometer was used to record the measurements. The enzyme solutions were prepared at 2.5 units/mL concentrationin 1% gelatin solution. BChE/AChE and compound solutions (50 μL each) in 2% DMSO at 10^-1-10^-6 mM concentration were added to 3 mL phosphate buffer (pH 8 &plusnm; 0.1)followed by incubation for 5 min at 25 °C. The reaction was commenced by the addition of acetylthiocholine iodide (ATC) (10 μL) and DTNB (50 μL) to the enzyme-inhibitor mixture. The yellow anion production was measured at 412 nm for 10 min. Using similar method, same enzyme solution without the inhibitor was processed which served as a control. The blank reading contained 10 μL substrate, 50 μL DTNB, 50 μL (2%) DMSO, and 3 mL buffer. The reaction was performed in triplicate. |
| Affinity data for this assay | |
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