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Assay Method Information

Assay Name:  Acetylcholinesterase Inhibitory Assay
Description:  Briefly, 140 μL of sodium phosphate buffer (pH 8.0), 20 μL of each tested compound with different concentrations (4, 20, and 100 μM) and 20 μL enzyme solution were mixed and incubated at room temperature for 15 min. The reactions were initiated by the addition of 10 μL of 0.01 M 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and 10 μL of 0.075 ATCI. The reaction solution was incubated at 37 C for 20 min, then the production of thiocholine reacts with DTNB to produce a yellow compound (5-thio-nitrobenzoate), which was detected at 412 nm. All tested samples and the positive control (berberine) [Nguyen et al., Bioorg. Med. Chem., 23:3126-3134] were dissolved in 10% analytical grade dimethyl sulfoxide (DMSO). The reaction was independently experimented for three times and recorded in 96-well microplates using a microplate reader (VersaMax).
Affinity data for this assay
 

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Last update November 1, 2007
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