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However, for NET assays, 100 nM GBR12935 and citalopram had been added into the sucrose Option to block [3H]MPP+ uptake by DAT and SERT. For SERT assays, a hundred nM GBR12935 and nomifensine were included to sucrose to dam [3H]5-HT uptake by DAT and Web. Synaptosomes have been preloaded with radiolabeled substrate in Krebs-phosphate buffer for one h to achieve constant state. Launch assays were initiated by addition of 850 μL of preloaded synaptosomes to check tubes that contains a hundred and fifty μL buffer Alternative with or devoid of test medicines. Incubations lasted thirty min for DAT and NET assays, and 5 min for SERT assays. The discharge response was terminated by speedy vacuum filtration, accompanied by quantitative resolve of radioactivity retained on filters by liquid scintillation counting.
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