| Assay Method Information | |
| | Inhibitory Activity of Human Recombinant SSAO/VAP-1 |
| Description: | Test purpose: The following method is used to determine the inhibitory activity of the compounds in the examples of the present invention on human recombinant SSAO/VAP-1.Test Materials:Human recombinant SSAO/VAP-1 (VAP-1, human) was purchased from Sigma, Cat. No. SRP6241;Amplex® Red Monoamine Oxidase Assay Kit was purchased from Invitrogen, Cat. No. A12214;384-well plate was purchased from Corning, Cat. No. 6005174;Amplex® Red Hydrogen PeroxidePeroxidase Assay Kit was purchased from Invitrogen, Cat. No. A22188.Benzylamine hydrochloride was purchased from Sigma, Cat. No. B5136-25G;DMSO (Dimethyl Sulfoxide) was purchased from Sigma, Cat. No. D2650-100ML;Test Method:The test compound was dissolved in DMSO and diluted 4 times to a total of 10 concentrations. In a 384-well plate, 25 μL of human recombinant SSAO/VAP-1 (1.6 μg/mL) was added into each well. 100 nL of test compounds at different concentrations were added to each well containing human recombinant SSAO/VAP-1, and the plate was incubated at room temperature for 30 min. After incubating for 30 min, 25 μL of Amplex® Red Monoamine Oxidase Assay Kit (a reaction mixture containing 200 μM Amplex Red reagent, 1 U/mL HRP and 1 mM benzylamine hydrochloride) was added into the corresponding wells, and the plate was incubated at room temperature in the dark for 60 min. After 60 min, PerkinElmer's Envision was used to read the fluorescence value (RFU) under excitation at 530-560 nm and emission at 590 nm. |
| Affinity data for this assay | |
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