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| | In vitro Binding Affinity Assay |
| Description: | The assay was performed in accordance with the method described by Owens et al. with slight modifications. Rat cerebral cortex was homogenized in 30 volumes of ice-cold 50mM Tris-HCl containing 150mM NaCl and 5mM KCl, pH 7.7, at 25°C and centrifuged at 20,000 x g for 20 min. Pellet was resuspended in 30 volumes of buffer and centrifuged again for 2 more times. The mixture of 240uL of the tissue suspension, 30uL of 1µM imipramine, 30uL of 1nM [3H]-citalopram and 100µL of the analyzed compound (10^-10 to 10^-4M) were incubated at 22°C for 1 h. Incubations were terminated by vacuum filtration and washed twice with 100µL of ice-cold buffer. The radioactivity was measured using a WALLAC 1409 DSA liquid scintillation counter. |
| Affinity data for this assay | |
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