| Assay Method Information | |
| | Binding Assay |
| Description: | For binding to the human A1AR, [3H]R-PIA (2 nM) was incubated with membranes (40 μg/tube) from CHO cells stably expressing the human A1AR at 25° C. for 60 min in 50 mM Tris.HCl buffer (pH 7.4; MgCl2, 10 mM) in a total assay volume of 200 μL. Nonspecific binding was determined using 10 μM of N6-cyclopentyladenosine. For human A2A AR binding, membranes (20 μg/tube) from HEK-293 cells stably expressing the human A2A AR were incubated with 15 nM [3H]CGS21680 at 25° C. for 60 min in 200 μL of 50 mM Tris.HCl, pH 7.4, containing 10 mM MgCl2. N-5′-ethyluronamidoadenosine (10 μM) was used to define nonspecific binding. Reaction was terminated by filtration with GF/B filters. |
| Affinity data for this assay | |
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