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Assay Method Information

Assay Name:  5-HT6 Binding Assay
Description:  Radioligand binding assays were performed using membranes from HEK-293 transfected with human 5-HT6 receptor. In these membranes the receptor concentration is 2.18 pmol/mg proteins and the protein concentration is 9.17 mg/mL. The incubation was initiated by addition of membrane to reaction buffer contained radioligand in the presence of test compound. And the incubation time was 60 min at 37 deg C. After incubation, the membranes were collected onto polyethylenimine-pretreated glass fiber filters. Radioactivity was determined by scintillation counting. Nonspecific binding was determined with 100 uM serotonin. Competition binding data were analyzed by using the LIGAND program, and assays were performed in triplicate determinations for each point. A linear regression line of data points is plotted, from which the concentration of competing ligand which displaces 50% of the specific binding of the radioligand (IC50 value) is determined and the Ki value is determined based upon the Cheng-Prusof equation. Ki was calculated when inhibition at 100 nM > 70%.
Affinity data for this assay
 

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Last update November 1, 2007
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