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In Situ Protein Binding Assay Using Fc-Fusion Proteins

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Synapse Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1538))

Abstract

This protocol describes an in situ protein-protein interaction assay between tagged recombinant proteins and cell-surface expressed synaptic proteins. The assay is arguably more sensitive than other traditional protein binding assays such as co-immunoprecipitation and pull-downs and provides a visual readout for binding. This assay has been widely used to determine the dissociation constant of binding of trans-synaptic adhesion proteins. The step-wise description in the protocol should facilitate the adoption of this method in other laboratories.

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Author information

Authors and Affiliations

  1. Department of Physiology and Pathophysiology, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada

    Nirmala Padmanabhan & Tabrez J. Siddiqui

  2. Neuroscience Research Program, Kleysen Institute for Advanced Medicine, Health Sciences Centre, University of Manitoba, 710 William Avenue, Winnipeg, MB, Canada, R3E 0Z3

    Nirmala Padmanabhan & Tabrez J. Siddiqui

Authors
  1. Nirmala Padmanabhan
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  2. Tabrez J. Siddiqui
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Corresponding author

Correspondence to Tabrez J. Siddiqui .

Editor information

Editors and Affiliations

  1. Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA

    Alexandros Poulopoulos

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Padmanabhan, N., Siddiqui, T.J. (2017). In Situ Protein Binding Assay Using Fc-Fusion Proteins. In: Poulopoulos, A. (eds) Synapse Development. Methods in Molecular Biology, vol 1538. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6688-2_7

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  • DOI: https://doi.org/10.1007/978-1-4939-6688-2_7

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6686-8

  • Online ISBN: 978-1-4939-6688-2

  • eBook Packages: Springer Protocols

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