| Assay Method Information | |
| | Binding affinity against monoamine oxidase-B (MAO-B) |
| Description: | MAO-B (M7441) was purchased from Sigma Aldrich Co. LLC (St. Louis, MO). The MAO-B membrane fractions (0.5 μg), and 0.1 to 10,000 nM of [3H] THK-5351 (1 μM) were mixed (200 μL), and the mixture was reacted at room temperature for 2 hours. The reaction solution was transferred to MultiScreen HTS 96-well 0.65 μL filtration plate (Millipore, Billerica, MA), and a B/F separation was conducted with a MultiScreen HTS Vacuum Manifold, and the mixture was washed with Dulbecco's PBS, and 0.1% BSA three times, and the filter was incubated with 2 mL of Scintillation fluid (Emulssifer-Safe; Perkin Elemer, Boston, MA), and a radio activity was measured with a beta counter (LS6500 liquid scintillation counter, Beckman Counter, Brea, CA). IC50 value was calculated by using Graph Pad Prism Version 7 (GraphPad Software, San Diego, CA). |
| Affinity data for this assay | |
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