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Single-Cell Imaging of ERK Signaling Using Fluorescent Biosensors

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Kinase Signaling Networks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1636))

Abstract

Single-cell analysis of the mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase (ERK) provides a means to perform highly detailed kinetic studies, assess heterogeneity between cells, and distinguish the subcellular localization of ERK activity. We describe here the methods needed to perform such measurements in a cell type of the investigator’s choosing. We discuss the selection of appropriate reporters and provide detailed methods for stably introducing reporters, collecting live-cell data, and automatically extracting quantitative information from individual cells.

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Acknowledgments

We thank Didem Sarikaya for helpful feedback on the manuscript. The methods described here were developed in part through support from the American Cancer Society (IRG-95-125-16).

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Authors and Affiliations

  1. Department of Molecular and Cellular Biology, University of California, Davis, 243 Briggs Hall, One Shields Avenue, Davis, CA, 95616, USA

    Michael Pargett, Taryn E. Gillies, Carolyn K. Teragawa, Breanne Sparta & John G. Albeck

Authors
  1. Michael Pargett
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  2. Taryn E. Gillies
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  3. Carolyn K. Teragawa
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  4. Breanne Sparta
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  5. John G. Albeck
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Corresponding author

Correspondence to John G. Albeck .

Editor information

Editors and Affiliations

  1. Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA

    Aik-Choon Tan

  2. Protein Networks Laboratory, Institute of Cancer Research, London, United Kingdom

    Paul H. Huang

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Pargett, M., Gillies, T.E., Teragawa, C.K., Sparta, B., Albeck, J.G. (2017). Single-Cell Imaging of ERK Signaling Using Fluorescent Biosensors. In: Tan, AC., Huang, P. (eds) Kinase Signaling Networks. Methods in Molecular Biology, vol 1636. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7154-1_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7154-1_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7152-7

  • Online ISBN: 978-1-4939-7154-1

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