Assay Method Information |
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| 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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