| Assay Method Information | |
| | Radioligand Binding Assay to HeLa Cells |
| Description: | After reaching confluence in 12-well or 24-well dishes, the cells were washed twice with ice-cold PBS and incubated for 2 hr with [3H]-AVP at 4 deg C. For the competition experiment, test compounds were dissolved in dimethyl sulfoxide, diluted with DMEM medium and added into the wells at several appropriate concentrations. After 2 hr of incubation, the cells were rinsed twice with ice-cold phosphate-buffered saline, lysed in 0.1 N NaOH containing 0.1% sodium dodecyl sulfate, transferred into scintillation vials, and detected using a liquid scintillation counter (LSC-1050, Aloka, Tokyo, Japan). The protein content of the lysed cells was determined by the method of Bradford. |
| Affinity data for this assay | |
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