| Assay Method Information | |
| | ADORA2B Receptor Binding Assay |
| Description: | 100 μL of Adenosine A2B membrane stock was dispensed into individual wells of an assay plate. 1 μL of each compound (test compounds, high control compound, low control compound) was then transferred into individual wells, followed by 100 μL of radio-labeled ligand. Plates were then sealed, and incubated at room temperature for 1 hour. Unifilter-96 GF/C filter plates were pre-soaked with 50 μL of 0.5% BSA per well for at least 30 min at room temperature. When binding assays were completed, reaction mixtures were filtered through GF/C plates using a Perkin Elmer Filtermate Harvester, and then each plate was washed 4× with cold wash buffer. Filter plates were then dried for 1 hour at 50° C. After drying, the bottom of the filter plate wells was sealed using Perkin Elmer Unifilter-96 backing seal tape. 50 μL of Perkin Elmer Microscint 20 cocktail was then added, and the top of the filter plate was then sealed using Perkin Elmer TopSeal-A sealing film. 3H trapped on the filter was counted using a Perkin Elmer MicroBeta2 Reader. Data analysis was performed using GraphPad Prism 5 software, and the Inhibition [% Control] was calculated using the following equation: % Inh=(1−Background subtracted Assay value/Background subtracted HC value) *100. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |