Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells

Single-molecule fluorescence recovery after photobleaching (smFRAP) is a newly developed technique that combines single-molecule super-resolution microscopy and traditional FRAP microscopy. smFRAP enables researchers to measure the dynamics, spatial locations, and relative concentrations of proteins...

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Detalles Bibliográficos
Autores principales: Tingey, Mark, Li, Yichen, Yang, Weidong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111821/
https://www.ncbi.nlm.nih.gov/pubmed/34007970
http://dx.doi.org/10.1016/j.xpro.2021.100490
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author Tingey, Mark
Li, Yichen
Yang, Weidong
author_facet Tingey, Mark
Li, Yichen
Yang, Weidong
author_sort Tingey, Mark
collection PubMed
description Single-molecule fluorescence recovery after photobleaching (smFRAP) is a newly developed technique that combines single-molecule super-resolution microscopy and traditional FRAP microscopy. smFRAP enables researchers to measure the dynamics, spatial locations, and relative concentrations of proteins. Here, we describe a step-by-step protocol for smFRAP on nuclear envelope transmembrane proteins on the inner nuclear membrane and outer nuclear membrane in live cells. For complete details on the use and execution of this protocol, please refer to Mudumbi et al. (2016a, 2016b, 2020 .
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spelling pubmed-81118212021-05-17 Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells Tingey, Mark Li, Yichen Yang, Weidong STAR Protoc Protocol Single-molecule fluorescence recovery after photobleaching (smFRAP) is a newly developed technique that combines single-molecule super-resolution microscopy and traditional FRAP microscopy. smFRAP enables researchers to measure the dynamics, spatial locations, and relative concentrations of proteins. Here, we describe a step-by-step protocol for smFRAP on nuclear envelope transmembrane proteins on the inner nuclear membrane and outer nuclear membrane in live cells. For complete details on the use and execution of this protocol, please refer to Mudumbi et al. (2016a, 2016b, 2020 . Elsevier 2021-04-30 /pmc/articles/PMC8111821/ /pubmed/34007970 http://dx.doi.org/10.1016/j.xpro.2021.100490 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Tingey, Mark
Li, Yichen
Yang, Weidong
Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
title Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
title_full Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
title_fullStr Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
title_full_unstemmed Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
title_short Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
title_sort protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8111821/
https://www.ncbi.nlm.nih.gov/pubmed/34007970
http://dx.doi.org/10.1016/j.xpro.2021.100490
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