| Assay Method Information | |
| | FRET Assay |
| Description: | Seed each cell lines (1×105 cells/well) into 96-well plates prior to experimentation.Incubate at 37° C. in 5% CO, for 24 hours.Wash each well with assay buffer (140 mM NaCl, 4:5 mM KCl, 10 mM D-Glucose, 2 mM CaCl2, 1 mM MgCl2, 10 mM HEPES, pH 7.4 adjusted with NaOH) twice.Add 1st loading solution containing 10 microM PTS18 and 0.06% Pluronic F-127 in assay buffer.Incubate the plate at rt in dark for 1 hour.Remove 1st loading solution and add 2nd loading solution containing 12.5 microM DiSBAC2(3), 1.25 mM Xylene Fast Yellow and 0.0075% Pluronic F-127 in assay buffer.Place the plate under the dark at rt for 25 minutes.Add compound solutions into the assay plate.Set the assay plate in FDSS and place an EFS device on the plate.Measure EFS-induced fluorescent response by FDSS. |
| Affinity data for this assay | |
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