Abstract
A goal of proteomic research is to detect all proteins in a particular biological subsystem, such as a cell type or cellular subfraction (1). This effort requires a combination of technologies first, for subfractionation and purification of the cells and cellular components of interest, and second, a sensitive method for detection and identification of all possible proteins. The challenge of proteomics is the wide range of abundance and sizes of cellular proteins, as well as the number of different proteins and posttranslational modifications. Clearly, maximizing both the specificity of the sample purification method and the sensitivity of the protein detection method is essential. In addition, targeting the analysis to specific sub-components of the cell can both enhance the sensitivity of the analysis and contribute to the functional analysis of the proteins (2).
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
Service, R. F. (2003) Public projects gear up to chart the protein landscape. Science 302(5649), 1316–1318.
Link, A. J., Eng, J, Schieltz, D. M., et al. (1999) Direct analysis of protein complexes using mass spectrometry. Nat. Biotechnol. 17(7), 676–682.
Shapiro H. (1994) Practical Flow Cytometry, Third Edition. Alan R. Liss, New York, NY.
Recktenwald, D. J., and Radbruch, A. (eds). (1998) Cell Separation Methods and Applications. Marcel Dekker, Inc., New York, NY.
Chelius, D., Huhmer, A. F., Shieh, C. H., et al. (2002) Analysis of the adenovirus type 5 proteome by liquid chromatography and tandem mass spectrometry methods. J. Proteome Res. 1(6), 501–13.
Chelius, D. and Bondarenko, P. V. (2002) Quantitative profiling of proteins in complex mixtures using liquid chromatography and mass spectrometry. J. Proteome Res. 1(4), 317–23.
Hovius, R., Lambrechts, H., Nicolay, K., and Kruijff, B. (1990) Improved methods to isolate and subfractionate rat liver mitochondria. Lipid composition of the inner and outer membrane. Biochim. Biophys. Acta 1021, 217–226.
Reinhart, P. H., Taylor, W. M., and Bygrave, F. L. (1982) A procedure for the rapid preparation of mitochondria from rat liver. Biochem. J. 204, 731–735.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Godfrey, W.L., Rudd, C.J., Iyer, S., Recktenwald, D. (2005). Purification of Cellular and Organelle Populations by Fluorescence-Activated Cell Sorting for Proteome Analysis. In: Walker, J.M. (eds) The Proteomics Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-890-0:067
Download citation
DOI: https://doi.org/10.1385/1-59259-890-0:067
Publisher Name: Humana Press
Print ISBN: 978-1-58829-343-5
Online ISBN: 978-1-59259-890-8
eBook Packages: Springer Protocols