CHO cells transfected with hAR subtypes were washed with phosphate-buffered saline, detached with tripsine and centrifuged for 10 min at 200× g. Cells were seeded in a 96-well white half-area microplate (Perkin Elmer, Boston, USA) in a stimulation buffer composed of Hank Balanced Salt Solution, 5 mM HEPES, 0.5 mM Ro 20–1724, 0.1% BSA. The cAMP levels were then quantified by using the AlphaScreen cAMP Detection Kit (Perkin Elmer, Boston, MA, USA), following the manufacturer’s instructions [54 (link)]. At the end of the experiments, the plates were read with a Perkin Elmer EnSight Multimode Plate Reader.
Free full text: Click here
