| Assay Method Information | |
| | MAO Assay |
| Description: | The capability of compounds described herein to inhibit MAO was determined using a modified Promega MAO-Glo assay kit. The substrate ((4S)-4,5-dihydro-2-(6-hydroxybenzothiazolyl)-4-thiazolecarboxylic acid) was converted by MAO to produce methyl ester luciferin. The luciferin was then converted by luciferase to produce oxyluciferin and light. The light was measured using a luminescence setting on the plate reader. A standard curve was prepared by serially diluting luciferin in reaction buffer. The standard kit reaction buffer was used 100 mM HEPES (pH 7.5), 5% glycerol. The substrate was dispensed into the wells so that the final concentration for MAO-A inhibition assay would be 10 μM and MAO-B inhibition assay would be 1 M. Inhibitors were diluted in DMSO to 4 and distributed to the appropriate wells. Clorgyline and Deprenyl served as positive controls. rhMAO-A and rhMAO-B were purchased from Sigma. MAO-A or MAO-B was added to the appropriate wells. The plate was incubated for 1 hour, at room temperature with gentle shaking (90 rpm). The detection agent was added at equal volume (48 μL detection agent to 48 μL reaction). The plate was incubated for 20 minutes, at room temperature with gentle shaking (90 rpm). The plate luminescence was read. Data processing was performed using Prism software. |
| Affinity data for this assay | |
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