| Assay Method Information | |
| | Cholinesterase Inhibition Assay |
| Description: | In a 96-well plate, 50 uL of the sample dissolved in phosphate buffer (8 mmol/L K2HPO4, 2.3 mmol/L NaH2PO4, 150 mmol/L NaCl, and 0.05% Tween 20 at pH 7.6) as well as 50 uL of the AChE/BChE solution (0.25 unit/mL), from Electrophorus electricus/Bovine serum in the same phosphate buffer, were added. The assay solutions, except the substrate, were pre-incubated with the enzyme for 30 min at room temperature. After pre-incubation the substrate was added. The substrate solution consists of Na2HPO4 (40 mmol/L), acetylthiocholine/butyrylthiocholine (0.24 mmol/L), and 5,5'-dithio-bis-(2-nitrobenzoic acid) (0.2 mmol/L, DTNB, Ellman's reagent). The absorbance of the yellow anion product, due to the spontaneous hydrolysis of substrate, was measured at 405 nm for 5 min on a Microtiter plate reader (Multiskan EX, Thermo, Vantaa, Finland). |
| Affinity data for this assay | |
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