| Assay Method Information | |
| | YFP-Halide Influx Assay for the CFTR-WT |
| Description: | For this purpose, HEK293 cells are transfected with plasmid DNA containing WT CFTR and seeded in 96 well plates (70,000 HEK cells/well). Two days after transfection, cells are treated with test compounds.The CFTR channels are activated by treatment with the cAMP inducer forskolin (10.67 μM) and a dose response of test compounds in 1×D-PBS (from Gibco, Cat n#14090-091) for 20 minutes prior to addition of an F solution (137 mM NaI, 2.7 mM KI, 1.76 mM KH2PO4, 10.1 mM Na2HPO4, 5 mM glucose). The I− induced quenching of fluorescence is recorded immediately after injection of for 7 seconds. The impact of a compound on the channel functionality can be measured by looking at the effect on fluorescence, and is expressed as (1-(fluorescence after 7 seconds (F)/fluorescence before injection (F0))) and an EC50 can be derived from a (1−F/F0) vs compound concentration plot. |
| Affinity data for this assay | |
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