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

Kitchen, Philip ; Salman, Mootaz M. ; Abir-Awan, Mohammed ; Al-Jubair, Tamim LU ; Törnroth- Horsefield, Susanna LU ; Conner, Alex C. and Bill, Roslyn M. (2020) In STAR Protocols 1(3).
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.... (More)

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).

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
STAR Protocols
volume
1
issue
3
article number
100157
publisher
Cell Press
external identifiers
  • pmid:33377051
  • scopus:85108258897
ISSN
2666-1667
DOI
10.1016/j.xpro.2020.100157
language
English
LU publication?
yes
id
73c42a50-f2a8-47eb-a606-6769f760c1fd
date added to LUP
2021-07-14 15:06:14
date last changed
2024-04-20 09:26:19
@article{73c42a50-f2a8-47eb-a606-6769f760c1fd,
  abstract     = {{<p>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).</p>}},
  author       = {{Kitchen, Philip and Salman, Mootaz M. and Abir-Awan, Mohammed and Al-Jubair, Tamim and Törnroth- Horsefield, Susanna and Conner, Alex C. and Bill, Roslyn M.}},
  issn         = {{2666-1667}},
  language     = {{eng}},
  number       = {{3}},
  publisher    = {{Cell Press}},
  series       = {{STAR Protocols}},
  title        = {{Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells}},
  url          = {{http://dx.doi.org/10.1016/j.xpro.2020.100157}},
  doi          = {{10.1016/j.xpro.2020.100157}},
  volume       = {{1}},
  year         = {{2020}},
}