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2, and 100 μM 1-methyl-3-isobutylxanthine(IBMX). Incubation was started by addition of membrane suspension and test compounds. Following 20 min incubation at 37 °C, intracellular cAMP levels were measured by EIA (enzyme-immunoassay), and a compound showing inhibitory effects on 5-HT-stimulated cAMP accumulation was classified as an antagonist.
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