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Detection of Active Caspases During Apoptosis Using Fluorescent Activity-Based Probes

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Programmed Cell Death

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1419))

Abstract

Activity-based probes (ABPs) are reactive small molecules that covalently bind to active enzymes. When tagged with a fluorophore, ABPs serve as powerful tools to investigate enzymatic activity across a wide variety of applications. In this chapter, we provide detailed methods for using fluorescent ABPs to detect the activity of caspases during the onset of apoptosis in vitro. We describe how these probes can be used to biochemically profile caspase activity in vitro using fluorescent SDS-PAGE as well as their application to imaging protease activity in live animals and tissues.

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Author information

Authors and Affiliations

  1. Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC, Australia

    Laura E. Edgington-Mitchell

  2. Department of Pathology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA, USA

    Matthew Bogyo

  3. Department of Microbiology and Immunology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA, USA

    Matthew Bogyo

Authors
  1. Laura E. Edgington-Mitchell
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  2. Matthew Bogyo
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Corresponding author

Correspondence to Laura E. Edgington-Mitchell .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Genetics, La Trobe Inst for Mol Sci, La Trobe Univ, Bundoora, Victoria, Australia

    Hamsa Puthalakath

  2. Department of Biochemistry and Genetics, La Trobe Inst for Mol Sci, La Trobe Univ, Bundoora, Victoria, Australia

    Christine J. Hawkins

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© 2016 Springer Science+Business Media New York

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Edgington-Mitchell, L.E., Bogyo, M. (2016). Detection of Active Caspases During Apoptosis Using Fluorescent Activity-Based Probes. In: Puthalakath, H., Hawkins, C. (eds) Programmed Cell Death. Methods in Molecular Biology, vol 1419. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3581-9_3

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  • DOI: https://doi.org/10.1007/978-1-4939-3581-9_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3579-6

  • Online ISBN: 978-1-4939-3581-9

  • eBook Packages: Springer Protocols

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