Unit

UNIT 18.2 Analysis of Caspase Activation During Apoptosis

  1. Scott H. Kaufmann1,
  2. Timothy J. Kottke1,
  3. L. Miguel Martins2,
  4. Alexander J. Henzing3,
  5. William C. Earnshaw3

Published Online: 1 AUG 2001

DOI: 10.1002/0471143030.cb1802s11

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Kaufmann, S. H., Kottke, T. J., Martins, L. M., Henzing, A. J. and Earnshaw, W. C. 2001. Analysis of Caspase Activation During Apoptosis. Current Protocols in Cell Biology. 11:18.2:18.2.1–18.2.29.

Author Information

  1. 1

    Mayo Clinic, Rochester, Minnesota

  2. 2

    Imperial Cancer Research Fund, London, United Kingdom

  3. 3

    University of Edinburgh, Edinburgh, Scotland, United Kingdom

Publication History

  1. Published Online: 1 AUG 2001
  2. Published Print: JUL 2001

Abstract

This unit describes three methods for the detection of caspase activation as cells undergo apoptosis. Simple and relatively quantitative enzymatic assays are provided using suitable substrates. Because the various low-molecular-weight substrates available for these assays are not selective, however, the assays do not accurately distinguish between various caspases. Immunoblotting is described for following the activation of specific caspases. When coupled with subcellular fractionation, this method can provide large amounts of temporal and spatial information about caspase activation. Finally, affinity labeling protocols are provided for detecting active caspases in whole-cell lysates or subcellular fractions.