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125I-17 and were further incubated at 37 °C for 2, 15, 30, 45, 60 and up to 180 min. Cells were collected by filtering at each time point and washed two times with 200 μL of cold binding buffer containing the appropriate cations. The plates were then transferred to Packard Multiscreen adapter plates (Cat# 6005178) and 100 μL of MicroScint-20 was added to the wells. The radioactivity associated with the cells was measured in a Packard Topcount. Cells incubated with 1% dimethyl sulfoxide were used as positive control and cells incubated with 10 μM unlabeled 17 were served as the negative control (non-specific background). The amount of non-specific radioactivity associated with the negative control cells was subtracted from all observations. The amount of radioactivity associated with the positive control cell was equated to 0% bound receptor sites. Receptor occupancy (% bound) of test compound was determined from the positive cell control. The data is the mean of duplicates.
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2 in the presence or absence of TS2/16. The plates were then transferred to Packard Multiscreen adapter plates (Cat# 6005178) and 100 μL of MicroScint-20 was added to the wells. Radioactive counts were quantified in a Packard Topcount. Data analysis was performed using the MRLCalc algorithm.
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