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Basic Techniques to Study FcεRI Signaling in Mast Cells

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Mast Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1220))

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Abstract

Mast cells are the crucial effector cells for allergic reactions. They are activated through the aggregation of the high-affinity IgE receptor (FcεRI) with allergen and allergen-specific IgE. Tyrosine phosphorylation of FcεRI subunits and various signaling proteins is an initial triggering event, leading to the activation of several signaling pathways in mast cells. Much has been learned from analysis of mast cells derived from gene-targeted mice. Therefore, in this chapter we will first describe how to generate mast cells from mouse bone marrow cells and how to correct the genetic defect by retroviral transduction. Then we will describe how to assess early activation events by measuring several protein-tyrosine kinases (PTKs) and serine/threonine kinases (PS/TKs) such as Akt (protein kinase B), protein kinase C (PKC), and JNK. As signal transduction is highly dependent on protein-protein interactions, we will describe experimental details of co-immunoprecipitation methods that are used to confirm such interactions.

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Authors and Affiliations

  1. Division of Cell Biology, La Jolla Institute for Allergy and Immunology, 9420 Athena Circle, San Diego, CA, 92037, USA

    Yuko Kawakami & Toshiaki Kawakami M.D., Ph.D.

Authors
  1. Yuko Kawakami
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  2. Toshiaki Kawakami M.D., Ph.D.
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Corresponding author

Correspondence to Toshiaki Kawakami M.D., Ph.D. .

Editor information

Editors and Affiliations

  1. Department of Medical Genetics, The Biomedical Research Centre, University of British Columbia, Vancouver, British Columbia, Canada

    Michael R. Hughes

  2. Department of Medical Genetics, The Biomedical Research Centre, University of British Columbia, Vancouver, British Columbia, Canada

    Kelly M. McNagny

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© 2015 Springer Science+Business Media New York

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Kawakami, Y., Kawakami, T. (2015). Basic Techniques to Study FcεRI Signaling in Mast Cells. In: Hughes, M., McNagny, K. (eds) Mast Cells. Methods in Molecular Biology, vol 1220. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1568-2_13

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  • DOI: https://doi.org/10.1007/978-1-4939-1568-2_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1567-5

  • Online ISBN: 978-1-4939-1568-2

  • eBook Packages: Springer Protocols

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