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Apoptotic Caspase Activation and Activity

Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 414)

Summary

Caspases are central to the execution of apoptosis. Their proteolytic activity is responsible for the demise of cells in many physiological and pathological states. Great advances in understanding caspases have been made using recombinant caspase expression and enzymatic characterization. Assays to measure caspase activity in apoptotic cell extracts and the development of a reconstituted cell-free assay were also critical in establishing the hierarchy in the caspase activation cascade and comprehend how caspase-9 is activated by the apoptosome. More recently, new tools such as activity-based probes allowed us to detect caspase activation in their working environment providing readout of the system with minimal interference. This chapter describes some of the methods used by our group to study the activation mechanisms of caspases and their activity.

Key Words

Activation activity-based probe apoptosis caspase kinetics 
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Notes

Acknowledgments

We are grateful to the members of our laboratory who provided some of the results presented in this chapter and for carefully reviewing protocols.

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Copyright information

© Humana Press Inc. 2008

Authors and Affiliations

  1. 1.The Burnham Institute for Medical ResearchLa Jolla
  2. 2.Program in Cell Death and Apoptosis Research, The Burnham Institute for Medical ResearchLa Jolla

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