Detection of active caspases during apoptosis using fluorescent activity-based probes

Laura E Edgington-Mitchell, Matthew Bogyo

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

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.

Original languageEnglish
Title of host publicationProgrammed Cell Death: Methods and Protocols
EditorsHamsa Puthalakath, Christine J Hawkins
Place of PublicationNew York NY USA
PublisherSpringer
Pages27-39
Number of pages13
Volume1419
ISBN (Electronic)9781493935819
ISBN (Print)9781493935796
DOIs
Publication statusPublished - 1 Apr 2016

Publication series

NameMethods in Molecular Biology
Volume1419
ISSN (Print)10643745

Keywords

  • Activity-based probes
  • Apoptosis
  • Caspases
  • Fluorescent SDS-PAGE
  • Imaging
  • Proteases

Cite this

Edgington-Mitchell, L. E., & Bogyo, M. (2016). Detection of active caspases during apoptosis using fluorescent activity-based probes. In H. Puthalakath, & C. J. Hawkins (Eds.), Programmed Cell Death: Methods and Protocols (Vol. 1419, pp. 27-39). (Methods in Molecular Biology; Vol. 1419). New York NY USA: Springer. https://doi.org/10.1007/978-1-4939-3581-9_3
Edgington-Mitchell, Laura E ; Bogyo, Matthew. / Detection of active caspases during apoptosis using fluorescent activity-based probes. Programmed Cell Death: Methods and Protocols. editor / Hamsa Puthalakath ; Christine J Hawkins. Vol. 1419 New York NY USA : Springer, 2016. pp. 27-39 (Methods in Molecular Biology).
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Edgington-Mitchell, LE & Bogyo, M 2016, Detection of active caspases during apoptosis using fluorescent activity-based probes. in H Puthalakath & CJ Hawkins (eds), Programmed Cell Death: Methods and Protocols. vol. 1419, Methods in Molecular Biology, vol. 1419, Springer, New York NY USA, pp. 27-39. https://doi.org/10.1007/978-1-4939-3581-9_3

Detection of active caspases during apoptosis using fluorescent activity-based probes. / Edgington-Mitchell, Laura E; Bogyo, Matthew.

Programmed Cell Death: Methods and Protocols. ed. / Hamsa Puthalakath; Christine J Hawkins. Vol. 1419 New York NY USA : Springer, 2016. p. 27-39 (Methods in Molecular Biology; Vol. 1419).

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

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Edgington-Mitchell LE, Bogyo M. Detection of active caspases during apoptosis using fluorescent activity-based probes. In Puthalakath H, Hawkins CJ, editors, Programmed Cell Death: Methods and Protocols. Vol. 1419. New York NY USA: Springer. 2016. p. 27-39. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-3581-9_3