| Assay Method Information | |
| | Radioligand Binding Assay |
| Description: | In all radioligand binding experiments, the samples were incubated in a final volume of 0.1 ml of binding buffer for 60 minutes at 25° C. and then filtered over Unifilter plates (Perkin Elmer) pre-treated for 2 hours to limit tracer non-specific binding. Filters were washed five times with 0.5 ml of ice-cold washing buffer (50 mM Tris-HCl pH 7.4, 5 mM MgCl, 1 mM EDTA), and 50 μL of Microscint 20 (Perkin Elmer) were added to each filter. The plates were incubated 15 min at room temperature on an orbital shaker and then counted with a TopCount™ for 1 min/well.For A1 receptor, the assay was performed with [3H]-DPCPX and membranes from CHO-Kl cells transfected with human A1 receptor (Euroscreen FAST-001B).For A2A receptor, the assay was performed with [3H-NECA and membranes from HEK293 cells transfected with human A2A receptor (Euroscreen FAST-002B).For A2B receptor, the assay was carried with [3H]-DPCPX and membranes prepared from HEK293 cells transfected with human A2B receptor (Euroscreen FAST-003B).For A3 receptor, the assay was carried out with [125IJ-MECA and membranes from CHO-Kl cells transfected with human A3 receptor (Euroscreen FAST-004B). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |