Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells
(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).
(Less)
- author
- Kitchen, Philip ; Salman, Mootaz M. ; Abir-Awan, Mohammed ; Al-Jubair, Tamim LU ; Törnroth- Horsefield, Susanna LU ; Conner, Alex C. and Bill, Roslyn M.
- organization
- publishing date
- 2020
- type
- Contribution to journal
- publication status
- published
- subject
- in
- STAR Protocols
- volume
- 1
- issue
- 3
- article number
- 100157
- publisher
- Cell Press
- external identifiers
-
- scopus:85108258897
- pmid:33377051
- 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
- 2024-07-27 17:50:02
@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}}, }