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| | In-vitro Monoamine Oxidase Inhibition Assay |
| Description: | The assay was performed in a polystyrene 96-well plate having flat bottom containing 200 μL of the total reaction mixture. For each assay, mixture was composed of 136 μL of 50 mM phosphate buffer (pH 7.4), 2 μL of test compound(0.1 mM) and 45 μL of enzyme pre-incubated at 37 °C for 10 min. The enzymatic reaction was initiated by the addition of 5 μL of the substrate (p-tyramine 3 mM) and 12 μL of Amplex Red reagent and further incubated at 37 °C for 10 min. Consequently the production of H2O2 and resorufin was measured using microplate fluorescence reader (FLx 800, Bio-Tek Instruments, Inc., Winooski, USA) based on the fluorescence produced at excitation and emission wavelength of 544 nm and 590 nm, respectively. For all the test compounds, assay was performed in triplicate. |
| Affinity data for this assay | |
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