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High-Resolution Imaging of Cellular Proteins pp 153–170Cite as

Bimolecular Fluorescence Complementation (BiFC) Analysis of Protein–Protein Interactions and Assessment of Subcellular Localization in Live Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1474))

Abstract

Bimolecular fluorescence complementation (BiFC) is a fluorescence imaging technique used to visualize protein–protein interactions (PPIs) in live cells and animals. One unique application of BiFC is to reveal subcellular localization of PPIs. The superior signal-to-noise ratio of BiFC in comparison with fluorescence resonance energy transfer or bioluminescence resonance energy transfer enables its wide applications. Here, we describe how confocal microscopy can be used to detect and quantify PPIs and their subcellular localization. We use basic leucine zipper transcription factor proteins as an example to provide a step-by-step BiFC protocol using a Nikon A1 confocal microscope and NIS-Elements imaging software. The protocol given below can be readily adapted for use with other confocal microscopes or imaging software.

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Author information

Authors and Affiliations

  1. Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, 575 Stadium Mall Drive, West Lafayette, IN, 47907, USA

    Evan P. S. Pratt, Jake L. Owens, Gregory H. Hockerman & Chang-Deng Hu

Authors
  1. Evan P. S. Pratt
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  2. Jake L. Owens
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  3. Gregory H. Hockerman
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  4. Chang-Deng Hu
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Corresponding author

Correspondence to Chang-Deng Hu .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, The University of Memphis, Memphis, Tennessee, USA

    Steven D. Schwartzbach

  2. Department of Biological Sciences, The University of Memphis, Memphis, Tennessee, USA

    Omar Skalli

  3. Department of Anatomy, Universidad Central del Caribe, Bayamon, Puerto Rico, USA

    Thomas Schikorski

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© 2016 Springer Science+Business Media New York

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Pratt, E.P.S., Owens, J.L., Hockerman, G.H., Hu, CD. (2016). Bimolecular Fluorescence Complementation (BiFC) Analysis of Protein–Protein Interactions and Assessment of Subcellular Localization in Live Cells. In: Schwartzbach, S., Skalli, O., Schikorski, T. (eds) High-Resolution Imaging of Cellular Proteins. Methods in Molecular Biology, vol 1474. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6352-2_9

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  • DOI: https://doi.org/10.1007/978-1-4939-6352-2_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6350-8

  • Online ISBN: 978-1-4939-6352-2

  • eBook Packages: Springer Protocols

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