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Detecting Caspase Activity in Drosophila Larval Imaginal Discs

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Caspases,Paracaspases, and Metacaspases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1133))

Abstract

Caspases are a highly specialized class of cell death proteases. Since they are synthesized as inactive full-length zymogens, activation—at least of effector caspases and to some extent also of initiator caspases—requires a proteolytic cleavage event, generating a large and a small subunit, two of each forming the active caspase. The proteolytic cleavage event generates neo-epitopes at both the C-terminus of the large subunit and the N-terminus of the small subunit. The cleaved Caspase-3 (CC3) antibody was raised against the neo-epitope of the large subunit and thus detects only cleaved, but not full-length, Caspase-3. Although raised against human cleaved Caspase-3, the CC3 antibody cross-reacts in other species and detects cleaved caspases, most notably DrICE and Dcp-1, in Drosophila. This protocol describes the procedure for use of the CC3 antibody to detect caspase activity in larval imaginal discs in Drosophila.

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Acknowledgement

We would like to thank Yun Fan, Ernesto Perez, and Jillian Lindblad for their technical expertise and review of the manuscript. This work was supported by grants from the National Institutes of Health (GM068016 and GM107789).

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Authors and Affiliations

  1. Department of Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA

    Caitlin E. Fogarty & Andreas Bergmann

Authors
  1. Caitlin E. Fogarty
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  2. Andreas Bergmann
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Editor information

Editors and Affiliations

  1. Dept of Plant Biology & Forest Genetics, Swedish Univ of Agricultural Sciences Linnean Center for Plant Biology, Uppsala, Sweden

    Peter V. Bozhkov

  2. Sanford-Burnham Medical Research Inst. Apoptosis & Cell Death Research Prgm, La Jolla, California, USA

    Guy Salvesen

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Fogarty, C.E., Bergmann, A. (2014). Detecting Caspase Activity in Drosophila Larval Imaginal Discs. In: V. Bozhkov, P., Salvesen, G. (eds) Caspases,Paracaspases, and Metacaspases. Methods in Molecular Biology, vol 1133. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0357-3_7

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  • DOI: https://doi.org/10.1007/978-1-4939-0357-3_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0356-6

  • Online ISBN: 978-1-4939-0357-3

  • eBook Packages: Springer Protocols

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