| Assay Method Information | |
| | Radioligand Binding Assays |
| Description: | For the A2a receptor binding study, the incubation contained 10 mg of the original tissue weight of the striatal membranes, 4 nM [3H]NECA, 50 nM CPA, 10 mM MgCl2, 0.2 units/mL adenosine deaminase, the test compound and 1% DMSO. For the A1 receptor binding study, the incubations contained 5 mg of the original tissue weight of the whole brain membranes, 0.1 nM [3H]DPCPX, 0.2 units/mL adenosine deaminase, the test compound and 1% DMSO. The incubations were vortexed and incubated for 60 min at 25 °C in a shaking waterbath. Half an hour after incubation was started, the incubations were vortexed again. The incubations were terminated via filtration. The tubes were washed twice with 4 mL ice-cold Tris buffer and the filters were washed once more with 4mL ice-cold Tris buffer. The damp filters were place in scintillation vials and 4 mL of scintillation fluid was added. The vials were shaken and incubated for two hours before being counted. |
| Affinity data for this assay | |
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