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From fixed to FRAP: measuring protein mobility and activity in living cells

Nature Cell Biology volume 3, pages E145–E147 (2001)Cite this article

Abstract

Experiments with fluorescence recovery after photobleaching (FRAP) started 30 years ago to visualize the lateral mobility and dynamics of fluorescent proteins in living cells. Its popularity increased when non-invasive fluorescent tagging became possible with the green fluorescent protein (GFP). Many researchers use GFP to study the localization of fusion proteins in fixed or living cells, but the same fluorescent proteins can also be used to study protein mobility in living cells. Here we review the potential of FRAP to study protein dynamics and activity within a single living cell. These measurements can be made with most standard confocal laser-scanning microscopes equipped with photobleaching protocols.

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Figure 1: Fluorescence recovery after photobleaching (FRAP).

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Acknowledgements

We thank A. Griekspoor for the illustration, and A. Benham, K. Jalink and C. Vos for useful comments on the manuscript. Our work is supported by a Pioneer grant from NWO, The Netherlands.

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Authors and Affiliations

  1. Division of Tumor Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    Eric A.J. Reits & Jacques J. Neefjes

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  1. Eric A.J. Reits
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  2. Jacques J. Neefjes
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Reits, E., Neefjes, J. From fixed to FRAP: measuring protein mobility and activity in living cells. Nat Cell Biol 3, E145–E147 (2001). https://doi.org/10.1038/35078615

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