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
Descripción
Sumario: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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