Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells

Philip Kitchen, Mootaz M. Salman, Mohammed Abir-Awan, Tamim Al-Jubair, Susanna Törnroth- Horsefield, Alex C. Conner, Roslyn M. Bill

Research output: Contribution to journalArticlepeer-review

1 Citation (SciVal)

Abstract

Aquaporins (AQPs) are membrane channel proteins that facilitate the movement of water down osmotic gradients across biological membranes. This protocol allows measurements of AQP-mediated water transport across the plasma membrane of live mammalian cells. Calcein is a fluorescent dye that is quenched in a concentration-dependent manner. Therefore, on short timescales, its concentration-dependent fluorescence can be used as a probe of cell volume, and therefore a probe of water transport into or out of cells. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Kitchen and Conner (2015). For the underlying methodology development, please refer to Fenton et al. (2010) and Solenov et al. (2004).

Original languageEnglish
Article number100157
JournalSTAR Protocols
Volume1
Issue number3
DOIs
Publication statusPublished - 2020

Subject classification (UKÄ)

  • Biochemistry and Molecular Biology

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