Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells
Summary: 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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721001970 |
| _version_ | 1819098364853092352 |
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| author | Mark Tingey Yichen Li Weidong Yang |
| author_facet | Mark Tingey Yichen Li Weidong Yang |
| author_sort | Mark Tingey |
| collection | DOAJ |
| description | Summary: 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. |
| first_indexed | 2024-12-22T00:29:49Z |
| format | Article |
| id | doaj.art-a0365e73a53641ada0e9cee32f4ddc3b |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-22T00:29:49Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-a0365e73a53641ada0e9cee32f4ddc3b2022-12-21T18:44:57ZengElsevierSTAR Protocols2666-16672021-06-0122100490Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cellsMark Tingey0Yichen Li1Weidong Yang2Department of Biology Temple University, Philadelphia, PA 19122, USADepartment of Biology Temple University, Philadelphia, PA 19122, USADepartment of Biology Temple University, Philadelphia, PA 19122, USA; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166721001970Atomic Force Microscopy (AFM)MicrobiologyMolecular BiologyBiotechnology and bioengineering |
| spellingShingle | Mark Tingey Yichen Li Weidong Yang Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and spatial locations of nuclear transmembrane proteins in live cells STAR Protocols Atomic Force Microscopy (AFM) Microbiology Molecular Biology Biotechnology and bioengineering |
| 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 | Atomic Force Microscopy (AFM) Microbiology Molecular Biology Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721001970 |
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