Abstract
Caspases are critical regulators of the apoptotic program, responsible for the harmonic dismantling of the cell. Cell death can occur by way of different options (necroptosis, necrosis, extreme autophagy) but once caspases are fully engaged it will take the apoptotic route. Hence, in general, caspase activity is inversely proportional to cell viability. Caspase activation can be measured by means of different techniques. Here, we describe three different methodologies to measure/observe caspase activation. Two approaches are recommended for studies on the whole cell population, whereas the third one is designed for visualizing caspase activation in single live cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Caroline, H. Y., and Yuan, J. (2009) The Jekyll and Hyde Functions of Caspases Dev. Cell 16, 21–34
Zong W.X., and Thompson, C.B. (2006) Necrotic death as a cell fate. Genes Dev. 20, 1–15
Pop, C., and Salvesen, G.S. (2009) Human caspases: activation, specificity, and regulation. J Biol Chem. 284, 21777–81
Paroni, G., Henderson, C., Schneider, C., and Brancolini, C. (2001) Caspase-2-induced apoptosis is dependent on caspase-9, but its processing during UV- or tumor necrosis factor-dependent cell death requires caspase-3. J Biol Chem. 276, 21907–1
Rodriguez J, Lazebnik Y. (1999) Caspase-9 and APAF-1 form an active holoenzyme. Genes Dev. 13, 3179–84
Henderson, C.J., Aleo, E., Fontanini, A., Maestro, R., Paroni, G., and Brancolini C. (2005) Caspase activation and apoptosis in response to proteasome inhibitors. Cell Death Differ. 12, 1240–54
Lüthi, A.U., and Martin, S.J. (2007) The CASBAH: a searchable database of caspase substrates. Cell Death Differ. 14, 641–50
Garcia-Calvo, M., Peterson, E.P., Leiting, B., Ruel, R., Nicholson, D.W., and Thornberry, N.A. (1998) Inhibition of human caspases by peptide-based and macromolecular inhibitors. J Biol Chem. 273, 32608–13
Rehm, M., Dussmann, H., Janicke, R.U., Tavare, J.M., Kogel, D., Prehn, J.H. (2002). Single-cell fluorescence resonance energy transfer analysis demonstrates that caspase activation during apoptosis is a rapid process. Role of caspase-3. J Biol Chem. 277, 24506–14
Acknowledgments
This work is supported by AIRC (Associazione Italiana Ricerca sul Cancro), MIUR (Ministero dell’Istruzione, Università e Ricerca), and Regione Friuli-Venezia Giulia (AITT-LR25/07).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Paroni, G., Brancolini, C. (2011). Measurement of Caspase Activity: From Cell Populations to Individual Cells. In: Stoddart, M. (eds) Mammalian Cell Viability. Methods in Molecular Biology, vol 740. Humana Press. https://doi.org/10.1007/978-1-61779-108-6_9
Download citation
DOI: https://doi.org/10.1007/978-1-61779-108-6_9
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-107-9
Online ISBN: 978-1-61779-108-6
eBook Packages: Springer Protocols