The AChE inhibition assay was described in our previous study.21 (link)
Briefly, recombinant human AChE (50 mU/mL) was dispensed (4 µL/well) into black clear-bottom 1536-well plates. Chlorpyrifos-oxon and BW284C51, known nonselective and selective AChE inhibitors, respectively, were used as positive controls. Twenty-three nanoliters of test compounds with concentrations ranging from 0.37 nM to 28.75 μM and controls were transferred into the assay plates using a Wako Pintool. After a 30 min incubation period at room temperature, 4 µL of colorimetric detection cocktail solution (DTNB, acetylthiocholine) was added to each well using a BioRaptr FRD (Beckman Coulter). The final DMSO concentration in the assay well was 0.29%. Assay plates were incubated at room temperature for another 10 min, followed by measuring the absorbance readout (405 nm) using an Envision plate reader (PerkinElmer).
Briefly, recombinant human AChE (50 mU/mL) was dispensed (4 µL/well) into black clear-bottom 1536-well plates. Chlorpyrifos-oxon and BW284C51, known nonselective and selective AChE inhibitors, respectively, were used as positive controls. Twenty-three nanoliters of test compounds with concentrations ranging from 0.37 nM to 28.75 μM and controls were transferred into the assay plates using a Wako Pintool. After a 30 min incubation period at room temperature, 4 µL of colorimetric detection cocktail solution (DTNB, acetylthiocholine) was added to each well using a BioRaptr FRD (Beckman Coulter). The final DMSO concentration in the assay well was 0.29%. Assay plates were incubated at room temperature for another 10 min, followed by measuring the absorbance readout (405 nm) using an Envision plate reader (PerkinElmer).
