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Assay Method Information

Assay Name:  In Vitro Activity Assay
Description:  Amine oxidase activity of recombinant SSAO, MAOa, and MAOb isoforms are measured using the MAO-Glo™ assay kit from Promega (V1402). Test compounds (with DMSO as vehicle, 0.5% v/v for SSAO) and the enzyme are incubated for 10 mins at room temperature before the addition of the luminogenic substrate. The substrate concentration is 10 μM for human recombinant SSAO. The assays are conducted in a pH 7.4 buffer (50 mM HEPES, 120 mM NaCl, 5 mM KCl, 2 mM CaCl2, 1.4 mM MgCl2, 0.001% Tween-20) in a well-plate. Oxidation of the substrate is conducted for 2 hrs before the addition of detecting reagent according the manufacture's protocol. The IC50 value of the tested compounds is calculated by fitting the dose response curve using a 4-parameter non-linear regression routine.
Affinity data for this assay
 

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Last update November 1, 2007
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