| Assay Method Information | |
| | Inhibition of DAO (Diamine Oxidase) |
| Description: | Test purpose: The following method is used to determine the selective inhibitory activity of the compound of the present invention on DAO.Test Materials:Human recombinant DAO (Recombinant Human ABP-1/DAO) was purchased from R&D, Cat. No. 8298-AO;Amplex® Red Hydrogen PeroxidePeroxidase Assay Kit was purchased from Invitrogen, Cat. No. A22188;1,4-Diaminobutane dihydrochloride was purchased from Aladdin, Cat. No. D106194-25G.Test Method:The test compound was dissolved in DMSO and diluted 5 times to a total of 6 concentrations. In a 384-well plate, 24 μL of human recombinant DAO (1 μg/mL) was added into each well. 1 μL of test compounds at different concentrations were added to each well containing human recombinant DAO, and the plate was incubated at 37° C. for 30 min. After incubating for 30 min, 25 μL of Amplex® Red Hydrogen Peroxide Peroxidase Assay Kit (containing 100 μM Amplex® Red and 0.2 U/ml HRP) containing 1 M 1,4-butanediamine dihydrochloride was added into the corresponding wells, and the plate was incubated in the dark at 37° C. for 30 min. After 30 min, PHERAstar FSX microplate reader of BMG LABTECH was used to read the fluorescence value (RFU) under excitation at 540 nm and emission at 580 nm. |
| Affinity data for this assay | |
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