| Assay Method Information | |
| | Binding Affinities to Different Adenosine Receptors |
| Description: | Binding affinity and specificalities of the compounds against different subtype of human adenosine receptors (hA1, hA2a, hA2b, and hA3) were characterized with cell member chromatography binding analysis. The compounds at different concentrations were incubate with hA1 membrane (from PerkinElmer) and [3H]-8-Cyclopenthyl-1-,3-dipropylxanthine (DPCPX) for 50 min at 25C, meanwhile 100uL 0.5% PEI solution was added into UNFILTER-96 GF/B filter palate for 60 min at 4C then UNFILTER-96 GF/B filter palate was washed twice with 50ml wash buffer, the membrane mix was transferred into UNFILTER-96 GF/B filter palate, and the filter plate was washed 4 times before incubated at 55C for 10 min. At last, 40 uL ULTIMA GOLD was added into each well, and CPM was read by TopCount. |
| Affinity data for this assay | |
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