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Assay Method Information

Assay Name:  AChE and BuChE Inhibition Assay
Description:  The reaction mixture was consisted of 100 無 of the total volume. To the solution of PBS (pH 8.0, 30 無) consisted of KH2PO4 and K2HPO4 in 96-well plate, the solution of enzyme AChE (15 無, 0.132 U/mL or BuChE (15 無, 0.04 U/mL) in the PBS was added and followed by the addition of the synthesized compounds (20 痠ol/L, 25 無 DMSO). The resulting mixture were thoroughly mixed, pre-incubated for 10 min at 37 蚓 and pre-read at 412 nm. The reaction started by the addition of 15 無 of 4mM well-1substrate (acetylthiocholine iodide-ATCI or butyrylthiocholine chloride-BuSCH), followed by the incorporation of 15 無 of 6 mM well-1 Ellman solution (DTNB). After 30 min of incubation at 37 蚓, absorbance was recorded at 412 nm by 96-well plate reader Bio-Rad Laboratories Inc. All experiments were performed with their respective standards (controls) in triplicate. As a positivecontrol Huperzine A (0.8 然 well-1) or Iso-OMPA (16.0 然 well-1) was practiced.
Affinity data for this assay
 

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Last update November 1, 2007
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