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Assay Method Information

Assay Name:  Acetylcholinesterase Inhibition Assay
Description:  AChI and 5,5'-dithio-bis(2-nitro-benzoic)acid (DTNB) were used as substrate for this enzymatic reaction. To this end, 100 L of Tris/HCl buffer (1 M, pH 8.0) and 10 L of novel bromophenols solution at different concentrations and 50 L AChE (5.32 10^-3 U) solution were mixed and incubated for 10 min at 25 C. Then 50 L of DTNB (0.5 mM) was transferred. Then the reaction was initiated by the addition of 50 L of AChI. The hydrolysis of AChI was recorded spectrophotometrically by the formation of yellow 5-thio-2-nitrobenzoate anion as the result of the reaction of DTNB with thiocholine at a wavelength of 412 nm.
Affinity data for this assay
 

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Last update November 1, 2007
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