| Assay Method Information | |
| | Binding Assay |
| Description: | CHO cells (ATCC No. CCL-61), in which A1 and A3 adenosine receptors were expressed, were cultured in F-12 media (Gibco, U.S.A.) supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 units/ml and 100 g/ml), at 37 °C in a 5% CO2 atmosphere. A predetermined amount of suitable hAR-expressed CHO cells was mixed with labeled ligands (1 nM [3H]CCPA and 0.5 nM [125I]AB-MECA) specifically binding to A1 and A3 adenosine receptors in a 50/10/1 buffer in test tubes . The derivatives of the present invention were dissolved at various concentrations in dimethylsulfoxide (DMSO) and diluted in the buffer, taking care that the final concentration of DMSO did not exceed 1%. Incubation for 1 hr in a 370C incubator was followed by rapid filtration in a vacuum using a cell collector (TOMTEC, U.S.A.). Subsequently, the test tubes were washed three times with 3 ml of the buffer before radioactivity was measured using a -counter. |
| Affinity data for this assay | |
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