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Caspases: determination of their activities in apoptotic cells

Alena Vaculova1, Boris Zhivotovsky

  • 1Institute of Environmental Medicine, Division of Toxicology, Karolinska Institute, Stockholm, Sweden.

Methods in Enzymology
|July 30, 2008
PubMed

Insights

Caspase enzymes are key regulators of programmed cell death (apoptosis) and have other roles in living cells. Detecting caspase activation is crucial for understanding biological processes and apoptosis, with various methods available for analysis.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Caspases, cysteine proteases, are central regulators of apoptosis.
  • They mediate key apoptotic events by cleaving numerous substrates.
  • Caspases also perform vital nonapoptotic functions in living cells.

Purpose of the Study:

  • To review methods for detecting and quantifying caspase activation.
  • To highlight the importance of caspase activity in biological processes.
  • To compare the advantages and disadvantages of various analytical techniques.

Main Methods:

  • Immunoblotting
  • Synthetic substrate cleavage assays
  • Affinity labeling
  • Flow cytometry
  • Microscopic techniques
  • In vivo caspase activity analysis

Main Results:

  • Caspase activity is a critical indicator of apoptosis.
  • Detection methods vary in sensitivity and application.
  • Both in vitro and in vivo approaches are discussed.

Conclusions:

  • Accurate detection of caspase activation is essential for studying apoptosis and other cellular functions.
  • A range of methods exists, each with specific strengths and limitations.
  • Understanding these methods aids in selecting appropriate techniques for research.

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