Abstract
Many single-molecule experimental techniques exploit fluorescence as a tool to investigate conformational dynamics, molecular interactions, or track the movement of proteins in order to gain insight into their biological functions. A prerequisite to these experimental approaches is to graft one or more fluorophores on the protein of interest with the desired photophysical properties. Here, we describe procedures for efficient methods used to covalently attach fluorophores to proteins. Alternative direct and indirect labeling strategies are also described.
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Acknowledgments
Work in our laboratory is supported by LASERLAB-EUROPE (grant agreement no 284464, EC’s Seventh Framework Programme), the ARC Foundation for Cancer Research and the French National Cancer Institute. We thank Sabrina Lignon, Marielle Bauzan, and Yann Denis of the Institut de Microbiologie de la Méditerranée technical platforms for advice and help with instrumentation and services. We thank Marc Wold (University of Iowa) for the gift of the p11d-tRPA polycistronic expression construct.
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Modesti, M. (2018). Fluorescent Labeling of Proteins. In: Peterman, E. (eds) Single Molecule Analysis. Methods in Molecular Biology, vol 1665. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7271-5_6
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DOI: https://doi.org/10.1007/978-1-4939-7271-5_6
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