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Et fluorescensbaseret assay af membranpotentiale til funktionel undersøgelse med høj kapacitet af to endogene ionkanaler i to epitelcellelinjer
JoVE Journal
Biology
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JoVE Journal Biology
A Fluorescence-Based Assay of Membrane Potential for High-Throughput Functional Study of Two Endogenous Ion Channels in Two Epithelial Cell Lines

Et fluorescensbaseret assay af membranpotentiale til funktionel undersøgelse med høj kapacitet af to endogene ionkanaler i to epitelcellelinjer

DOI:
10.3791/63528-v

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06:59 min

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June 22, 2022

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, , ,
1Molecular Medicine,Hospital for Sick Children, 2Cell Biology,Hospital for Sick Children, 3Department of Physiology,University of Toronto, 4Department of Paediatrics,University of Toronto, 5Department of Biochemistry,University of Toronto

Chapters

  • 00:05Introduction
  • 00:46Cell Plating and Differentiation
  • 02:33CFTR Functional Assay
  • 05:24Results: Measurement of CFTR Channel Activity
  • 06:22Conclusion

Summary

Automatic Translation

Automatic Translation

Denne protokol beskriver en metode til undersøgelse af elektrogene membranproteiner ved at måle ændringer i membranpotentiale. Dette assay giver en platform for funktionel aflæsning af flere ionkanaler, der endogent udtrykkes i epitelcellelinjer.

Tags

Fluorescence-based AssayMembrane PotentialHigh-throughputIon ChannelsEpithelial CellsCalu-3Caco-2Cell CultureTrypsin96-well Plate384-well PlateSodium-free BufferChloride-free BufferNMDG

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