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Cell Migration pp 387–402Cite as

Fluorescence Recovery After Photobleaching

Part of the Methods in Molecular Biology book series (MIMB,volume 769)

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.

Key words

  • Fluorescence recovery after photobleaching
  • Fluorescence microscopy
  • Green fluorescent protein
  • Half-time of recovery
  • Mobile fraction
  • Diffusion
  • Binding reaction kinetics
  • Focal adhesions
  • Vinculin

Alex Carisey, Matthew Stroud, and Ricky Tsang contributed equally to the manuscript.

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  • DOI: 10.1007/978-1-61779-207-6_26
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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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Authors and Affiliations

  1. Faculty of Life Sciences, The University of Manchester, Manchester, UK

    Christoph Ballestrem

Authors
  1. Alex Carisey
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  2. Matthew Stroud
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  3. Ricky Tsang
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  4. Christoph Ballestrem
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Corresponding author

Correspondence to Christoph Ballestrem .

Editor information

Editors and Affiliations

  1. , Division of Cancer Studies, King's College London, London, SE1 1UL, United Kingdom

    Claire M. Wells

  2. Randall Division of Cell and, Molecular Biophysics, King's College London, London, SE1 1UL, United Kingdom

    Maddy Parsons

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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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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-206-9

  • Online ISBN: 978-1-61779-207-6

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