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Assay Method Information

Assay Name:  DAAO in Vitro Activity Assay
Description:  Inhibitory effect of compounds was determined in a cell free fluorescence assay. The H2O2 generated from the degradation of D-serine was linked to oxidation of Amplex Red in the presence of horseradish peroxidase (HRP). Inhibitor compounds are diluted in half log increments to create an 11-point dose response curve. Each dilution is spotted in duplicate into black 384-well plates. The assay buffer containing DAAO, flavin adenine dinucleotide, horseradish peroxidase, and Amplex Red were added to each well of the plate using a Titertek MultiDrop-384 reagent addition device. Next, assay buffer containing D-serine was added using the MultiDrop. The reaction was then incubated in the dark for10-30 min before reading the plates on a Perkin-Elmer Envision 2103 multilabel reader using the following settings: 10 flashes of the flash lamp, excitation filter 530 nm, emission filter 590 nm. The nonlinear curve fitting was used to calculate an IC50 value for each compound.
Affinity data for this assay
 

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Last update November 1, 2007
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