Abstract
Since O’Farrell (1) introduced the improved technique for high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), it has become one of the most powerful tools for the separation and quantification of proteins from complex mixtures. The major reason for this success is that 2-D PAGE employs separation of denatured proteins according to two different parameters, mol wt and isoelectric point. Consequently, it has sufficient resolution to separate individual proteins as discrete spots on the gel. Each parameter may also be varied and therefore, with the modification of nonequilibrium pH-gradient electrophoresis (NEPHGE) to analyze basic proteins (2), almost any polypeptide may be investigated. Thus to date, the O’Farrell 2-D gel system has no serious rivals, with the possible exception of the Kaltschmidt and Wittmann (3) gel system for analyzing ribosomal proteins. Ribosomal proteins, however, may be adequately separated with NEPHGE.
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
O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021.
O’Farrell, P. Z., Goodman, H. M., and O’Farrell, P. H. (1977) High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12, 1133–1142.
Kaltschmidt, E. and Whittmann, H. G. (1970) Ribosomal proteins. VII. Two dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Ann. Biochem. 36, 401–412.
Celis, J. and Bravo, R. (eds.) (1983) Two Dimensional Gel Electrophoresis of Proteins. Academic, San Diego, CA.
Duncan, R. and McConkey, E. H. (1982) How many proteins are there in a typical mammalian cell? Clin. Chem. 28, 749–755.
Pollard, J. W. (1983) Application of two-dimensional polyacrylamide gel electrophoresis to studies of mistranslation in animal and bacterial cells, in Two-Dimensional Gel Electrophoresis of Proteins (Celis, J. and Bravo, R., eds.), Academic, San Diego, CA, pp. 363–395.
Parker, J., Pollard, J. W., Friesen, J. D., and Stanners, C. P. (1978) Stuttering: High level mistranslation in animal and bacterial cells. Proc. Natl. Acad. Sci. USA 75, 1091–1095.
Anderson, N. G. and Anderson, L. (1982) The human protein index. Clin. Chem. 28, 739–748.
Celis, J. E., Rasmussen, H. H., Leffers, H., Madsen, P., Honore, B., Gesser, B., Dejgaard, K., and Vandekerckhove, J. (1991) Human cellular protein patterns and their link to genome DNA sequence data: usefulness of two-dimensional gel electrophoresis and microsequencing. FASEB J. 5, 2200–2207.
Garrels, J. I. and Franza, B. R. (1989) The REF52 protein database. Methods of database construction and analysis using the Quest system and characterizations of protein patterns from proliferating and quiescent REF52 cells. J. Biol. Chem. 264, 5283–5298.
Special Issue: Two-dimensional gel electrophoresis in cell biology. (Celis, J. E., ed.), Electrophoresis 11 (1990).
Special Issue: Two-dimensional gel electrophoresis. Clin. Chem. 28 (1982).
Dunbar, B. S. (1988) Two-Dimensional Electrophoresis and Immunological Techniques. Plenum, New York.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1994 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Pollard, J.W. (1994). Two-Dimensional Polyacrylamide Gel Electrophoresis of Proteins. In: Walker, J.M. (eds) Basic Protein and Peptide Protocols. Methods in Molecular Biology™, vol 32. Humana Press. https://doi.org/10.1385/0-89603-268-X:73
Download citation
DOI: https://doi.org/10.1385/0-89603-268-X:73
Publisher Name: Humana Press
Print ISBN: 978-0-89603-268-2
Online ISBN: 978-1-59259-519-8
eBook Packages: Springer Protocols