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Protein Micropatterning Assay: Quantitative Analysis of Protein–Protein Interactions

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Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1550))

Abstract

Characterization, especially quantification, of protein interactions in live cells is usually not an easy endeavor. Here, we describe a straightforward method to identify and quantify the interaction of a membrane protein (“bait”) and a fluorescently labeled interaction partner (“prey”) (membrane-bound or cytosolic) in live cells using Total Internal Reflection Fluorescence microscopy. The bait protein is immobilized within patterns in the plasma membrane (e.g., via an antibody); the bait–prey interaction strength can be quantified by determining the prey bulk fluorescence intensity with respect to the bait patterns. This method is particularly suitable also for the analysis of weak, transient interactions that are not easily accessible with other methods.

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Acknowledgments

This work was funded by the Austrian Science Fund (FWF projects P 26337 and P 25730), the Austrian Research Promotion Agency (FFG project 842379), the program ‘Regionale Wettbewerbsfähigkeit OÖ 2007–2013’ with the financial support of the European Fund for Regional Development, as well as the Federal State of Upper Austria.

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Authors and Affiliations

  1. Institute of Applied Physics, TU Wien, Getreidemarkt 9, 1060, Vienna, Austria

    Gerhard J. Schütz & Eva Sevcsik

  2. School of Engineering and Environmental Sciences, University of Applied Sciences Upper Austria, Stelzhamerstrasse 23, 4600, Wels, Austria

    Julian Weghuber & Peter Lanzerstorfer

Authors
  1. Gerhard J. Schütz
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  2. Julian Weghuber
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  3. Peter Lanzerstorfer
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  4. Eva Sevcsik
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Corresponding author

Correspondence to Gerhard J. Schütz .

Editor information

Editors and Affiliations

  1. Keck School of Medicine, University of Southern California, Los Angeles, California, USA

    Lucio Comai

  2. Keck School of Medicine, University of Southern California, Los Angeles, California, USA

    Jonathan E. Katz

  3. Stanford University, Palo Alto, California, USA

    Parag Mallick

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Schütz, G.J., Weghuber, J., Lanzerstorfer, P., Sevcsik, E. (2017). Protein Micropatterning Assay: Quantitative Analysis of Protein–Protein Interactions. In: Comai, L., Katz, J., Mallick, P. (eds) Proteomics. Methods in Molecular Biology, vol 1550. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6747-6_18

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  • DOI: https://doi.org/10.1007/978-1-4939-6747-6_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6745-2

  • Online ISBN: 978-1-4939-6747-6

  • eBook Packages: Springer Protocols

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