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Ion Channels pp 65–77Cite as

Spectroscopy-Based Quantitative Fluorescence Resonance Energy Transfer Analysis

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 337))

Abstract

The combination of green fluorescent protein mutants and fluorescence resonance energy transfer (FRET) forms a powerful tool for ion channel studies. A key to successful application of green fluorescent protein-based FRET is to reliably separate the FRET signal from various non-FRET fluorescence emissions that coexist in any experimental system. This chapter introduces a FRET quantification method that is based on fluorescence spectroscopic microscopy. Application of this “spectra FRET” method to both the confocal imaging of Xenopus oocytes and the epifluorescence imaging of culture cells is described. The fluorescence intensity ratio measurement, a complementary non-FRET method for identifying the channel subunit stoichiometry, is also discussed.

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Author information

Authors and Affiliations

  1. Department of Physiology and Membrane Biology, University of California at Davis, School of Medicine, Davis, CA

    Jie Zheng

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  1. Jie Zheng
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Editor information

Editors and Affiliations

  1. University of Texas Health Science Center, San Antonio, TX, USA

    James D. Stockand  & Mark S. Shapiro  & 

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© 2006 Humana Press Inc.

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Zheng, J. (2006). Spectroscopy-Based Quantitative Fluorescence Resonance Energy Transfer Analysis. In: Stockand, J.D., Shapiro, M.S. (eds) Ion Channels. Methods in Molecular Biology™, vol 337. Humana Press. https://doi.org/10.1385/1-59745-095-2:65

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  • DOI: https://doi.org/10.1385/1-59745-095-2:65

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-576-7

  • Online ISBN: 978-1-59745-095-9

  • eBook Packages: Springer Protocols

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