Abstract
This chapter describes the use of microscope-based fluorescence recovery after photobleaching (FRAP). To quantify the dynamics of proteins within a subcellular compartment, we first outline the general aspects of FRAP experiments and then provide a detailed protocol of how to measure and analyse the most important parameters of FRAP experiments such as mobile fraction and half-time of recovery.
Alex Carisey, Matthew Stroud, and Ricky Tsang contributed equally to the manuscript.
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Acknowledgments
We would like to thank Dr. Janet Askari for critical reading of the manuscript. CB acknowledges BBSRC (BB/G004552/1). The Bioimaging Facility microscopes used in this study were purchased with grants from BBSRC, Wellcome Trust and the University of Manchester Strategic Fund.
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Carisey, A., Stroud, M., Tsang, R., Ballestrem, C. (2011). Fluorescence Recovery After Photobleaching. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology, vol 769. Humana Press. https://doi.org/10.1007/978-1-61779-207-6_26
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DOI: https://doi.org/10.1007/978-1-61779-207-6_26
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