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| | cAMP Production Assay |
| Description: | hese assays were performed at adenosine receptors transfected using a cAMP enzymeimmunoassay kit (Amersham Biosciences). CHO-A2A cells were seeded (10000 cells/well) in 96-well culture plates and incubated at 37 C. in an atmosphere with 5% CO2 in Dulbecco's Modified Eagle's Medium Nutrient Mixture F-12 (DMEM F-12), containing 10% Foetal Calf Serum (FCS) and 1% L-Glutamine. Cells were washed 3x with 200 ul assay medium (DMEM-F12 and 25 mM HEPES pH=7.4) and pre-incubated with assay medium containing 30 uM rolipram and test compounds at 37 C. for 15 min. 1 uM NECA was incubated for 15 min at 37 C. (total incubation time 30 min). Reaction was stopped with lysis buffer supplied in the kit and the enzymeimmunoassay was carried out for detection of intracellular cAMP at 450 nm in an Ultra Evolution detector (Tecan). |
| Affinity data for this assay | |
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