Abstract
As interest in cell death has recently increased, many new techniques have been developed to detect and quantitate apoptosis in cells. Multiparameter flow cytometric-based assays provide statistical information on measured parameters at the single-cell level, and are therefore well suited for measuring apoptosis (1). Since apoptosis has many functional expressions, it is often desirable for investigators to demonstrate cell death by several different methods. A sampling of flow cytometric assays used in our laboratory is presented here.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Darzynkiewicz, Z., Juan, G., Li, X., Gorczynca, W., Murakami, T, and Traganos, F (1997) Cytometry in cell necrobiology: anaysis of apoptosis and accidental cell death. Cytometry 27, 1–20.
Arends, M. J., Morris, R. G., and Wyllie, A. H. (1990) Apoptosis: the role of endonuclease. Am. J. Pathol. 136, 593–608.
Compton, M. M. (1992) A biochemical hallmark of apoptosis: internucleosomal degradation of the genome. Cancer Metast. Rev. 11, 105–119.
Wyllie, A. H. (1992) Apoptosis and the regulation of cell numbers in normal and neoplastc tissues. An overview. Cancer Metast. Rev. 11, 95–103.
Li, X. and Darzynkiewicz, Z. (1995) Labelling DNA strand breaks with BrdUTP. Detection of apoptosis and BrdUTP incorporation. Cell Prolif. 28, 572–579.
Kalejta, R. F., Shenl, T., and Beavis, A. S. (1997) Use of a membrane-localized green fluorescent protein allows simultaneous identification of transfected cells and cell cycle analysis by flow cytometry. Cytometry 29, 286–291.
Thornberry, N. A. and Lazebnik, Y. (1998) Caspases: enemies within. Science 291, 1312–1316.
Komoriya, A., Packard, B. Z., Brown, M. J., Wu, M.-L., and Henkart, P. A. (2000) Assesment of caspase activities in intact apoptotic thymocytes using cell-permeable fuorogenis caspase substrates. J. Exp. Med. 191, 1819–1828.
Fadok, V. A., VoelKer, D. R., Cammpbell, P. A., Cohen, J. J., Bratton, D. L., and Henson, P. M. (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148, 2207–2216.
Van Engeland, M., Nieland, L., Ramaekers, F., Schutte, B., and Reutelingsperger, C. (1998) Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry 31, 1–9.
Zhang, C., Ao, Z., Seth, A., and Schlossman, S. F. (1996) A mitochondrial membrane protein defined by a novel monoclonal antibody is preferentially detected in apoptotic cells. J. Immunol. 157, 3980–3987.
Petit, X., Lecouer, H., Zorn, E., Dauguet, C., Mignotte, B., and Gougeon, M.-L. (1997) Alterations in mitochondrial structure and function are early events of dexamethasone-induced thymocyte apoptosis. J. Cell Biol. 130, 157–167.
Marchetti, P., Decaudin, D., Macho, A., et al. (1997) Redox regulation of apoptosis: impact of thiol oxidation status on mitochondrial function. Eur. J. Immunol. 27, 289–296.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Dicker, D.T., El-Deiry, W.S. (2003). Flow Cytometric Analysis of Tumor Suppressor Gene-Induced Apoptosis. In: El-Deiry, W.S. (eds) Tumor Suppressor Genes. Methods in Molecular Biology™, vol 223. Humana Press. https://doi.org/10.1385/1-59259-329-1:271
Download citation
DOI: https://doi.org/10.1385/1-59259-329-1:271
Publisher Name: Humana Press
Print ISBN: 978-0-89603-987-2
Online ISBN: 978-1-59259-329-3
eBook Packages: Springer Protocols