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Cell Migration pp 387-402 | Cite as

Fluorescence Recovery After Photobleaching

  • Alex Carisey
  • Matthew Stroud
  • Ricky Tsang
  • Christoph BallestremEmail author
Protocol
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 

Notes

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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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Alex Carisey
  • Matthew Stroud
  • Ricky Tsang
  • Christoph Ballestrem
    • 1
    Email author
  1. 1.Faculty of Life SciencesThe University of ManchesterManchesterUK

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