| Assay Method Information | |
| | Uptake Assays |
| Description: | The uptake assays were carried out 2 days after transfection. Prior to the experiment, the cells were washed once in 500 ul of uptake buffer (25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 130 mM NaCl, 5.4 mM KCl, 1.2 mM CaCl2, 1.2 mM MgSO4, 1 mM L-ascorbic acid, 5 mM D-glucose, and 1 uM of the catechol-O-methyltransferase inhibitor Ro 41-0960 (Sigma), pH 7.4) at room temperature (RT). The unlabeled ligand (e.g. modafinil [(+-)-1] or analogues) was added to the cells in 10 concentrations from 1 nM to 0.1 mM equally distributed around the expected IC50 value, and uptake was initiated by addition of 10 nM radioligand in a final volume of 500 uL. |
| Affinity data for this assay | |
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