In vivo imaging and biochemical characterization of protease function using fluorescent activity-based probes

Laura E Edgington, Matthew Bogyo

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

14 Citations (Scopus)

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 article, detailed protocols are provided for using fluorescent ABPs to biochemically characterize the activity of proteases in vitro. Furthermore, descriptions are provided of how these probes can be applied to image protease activity in live animals and tissues along with subsequent analysis by histology, flow cytometry, and SDS-PAGE.

Original languageEnglish
Title of host publicationCurrent Protocols in Chemical Biology
PublisherJohn Wiley & Sons
Pages25-44
Number of pages20
Volume5
Edition1
ISBN (Electronic)9780470559277
DOIs
Publication statusPublished - 2013
Externally publishedYes

Keywords

  • activity‐based probes
  • caspases
  • imaging
  • proteases

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