Abstract
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) separates proteins by their iso-electric points (IEPs) and electro-phoretic mobilities (1). In the first dimension, a sample is applied to an iso-electric focusing (IEF) acrylamide gel. Within this the con stituent proteins migrate through a pH gradient until they reach their IEPs. A protein’s IEP is the pH value where it has no net charge. This is as a result of the positively and negatively charged amino acids cancelling each other. Proteins in a sample thus become focused at their individual IEPs. By the end of the electro-focusing there will be essentially no movement of proteins. An alternative to iso-electric focus ing is nonequilibrium pH gradient electrophoresis (NEPHGE) (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
O’Farrell, P H. (1975) High-resolution two dimensional electrophoresis of proteins. J Biol. Chem 250,4007–021.
O’Farrell, P Z., Goodman, H. M, and O’Farrell, P. H(1979) High resolution two dimensional electrophoresis of basic as well as acidic proteins Cell 12, 1133–1142.
Hames, B. D. (1981) An introduction to polyacrylamide gel electrophoresis, in Gel Electrophoresis of Proteins. A Practical Approach (Hames, B D. and Rickwood, D., eds.), IRL, London, pp. 1–9
Wellems, T. H and Howard, R. J (1986) Homologous genes encode two distinct histidine rich proteins. Proc. Natl. Acad. Sci. USA 83, 6065–6069.
Laskey, R. A and Mills, A. D. (1975) Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem 56, 335–341
Dunbar, B S. (1987) Two Dimensional Electrophoresis and Immunological Techniques. Plenum, New York.
Cells, J. and Bravo, R. (1984) Two Dimensional Gel Electrophoresis of Proteins. Methods and Applications. Academic, London.
Saravia, N. G, Gemmel, M A., Nance, S. L., and Anderson, N. L. (1984) Two-dimensional electrophoresis used to differentiate the causal agents of American tegumentary Leishmaniasis Clin Chem 30, 2048–2052.
Anderson, N. L., Parish, N. M., Richardson, J. P., and Pearson, T. W. (1985) Comparison of African trypanosomes of different antigenic phenotypes, subspecies and life cycle stages by two dimensional electrophoresis Mol. Biochem Parasitol. 16,299–314
Creasey, A. C, Fenton, B., Walker, A., Thaithong, S., Oliveira, S., Mutambu, S., and Walliker, D. (1990) Genetic diversity of Plasmodium falciparum shows geographical variation. Trans. Am. J. Trop. Med. Hyg. 42,403–413
Pinswasdi, C, Thaithong, S., Beale, G H., Fenton, B, Webster, H. K., and Pavanand, K. (1987) Polymorphism of proteins in malaria parasites following mefloquine treatment. Mol Biochem. Parasitol 23, 159–164
Wellems, T E., Oduola, A. M. J, Fenton, B., Desjardins, R., Panton, L. J., and Rosario, V. E. (1988) Chromosome size variation occurs in cloned Plasmodium falciparum on in vitro cultivation. Revista Brasil de Genetica 11,813–825.
Fenton, B. and Walliker, D (1990) Genetic analysis of polymorphic proteins of the human malaria parasite Plasmodium falciparum. Genet. Res. 55, 81–86.
Towbin, H., Staehelin, T, and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets, procedure and some applications. Proc. Natl. Acad. Sci. USA 76, 4350–4354.
Brown, G V, Anders, R. F., and Knowles, G. (1983) Differential effect of immunoglobulin on the in vitro growth of several isolates of Plasmodium falciparum Infect. Immun. 39, 1228–1235.
Fenton, B. Clark, J. T. Wilson, C. F., McBnde, J. S, and Walliker, D. (1989) Polymorphism of a 35-48 kDa Plasmodium falciparum merozoite surface antigen. Mol. Biochem Parasitol. 34, 79–86.
Fenton, B., Clark, J T, Robinson, J. V., Kahn, A., Walliker, D., Ridley, R, Scaife, J. G., and McBride, J. (1991) Structural and antigenic polymorphism of the 35-48 kilodalton merozoite surface antigen (MSA-2) of the malaria parasite Plasmodium falciparum. Mol. Cell Biol. 11,963–971.
Prioli, R. P, Mejia, J S, and Pereira, M. E A (1990) Monoclonal antibodies against Trypanosoma cruzi neuraminidase reveal enzyme polymorphism, recognize a subset of trypomastigotes, and enhance infection in vitro. J Immunol. 144,4384–4391.
Ouaissi, M. A, Taibi, A., Cornette, J., Velge, P., Marty, B, Loyens, M., Esteva, M., Rizvi, F. S., and Capron, A. (1990) Characterization of major surface and excretory-secretory immunogens of Trypanosoma cruzi trypomastigotes and identification of potential protective antigen. Parasitology 100, 115–124
Fenton, B. (1987) Peptide digest studies of polymorphic proteins of Plasmodium falciparum. Mol. Biochem Parasit. 24, 13–22
Neale, K. A., and Alderete, J. F. (1990) Analysis of proteinases of representative Trichomonas vaginalis isolates. Infect. Immun. 58, 157–162.
Fenton, B., Walker, A., and Walliker, D. (1985) Protein variation in clones of Plasmodium falciparum detected by two dimensional electrophoresis. Mol Biochem Parasitol 16, 173–183.
Sanderson, A, Walliker, D, and Molez, J-F. (1981) Enzyme typing of Plasmodium falciparum from some African and other old world countries. Trans R. Soc Trop. Med Hyg. 75, 263–267
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1993 Humana Press Inc, Totowa, NJ
About this protocol
Cite this protocol
Fenton, B. (1993). Two-Dimensional Polyacrylamide Gel Electrophoresis. In: Hyde, J.E. (eds) Protocols in Molecular Parasitology. Methods in Molecular Biology™, vol 21. Humana Press. https://doi.org/10.1385/0-89603-239-6:349
Download citation
DOI: https://doi.org/10.1385/0-89603-239-6:349
Publisher Name: Humana Press
Print ISBN: 978-0-89603-239-2
Online ISBN: 978-1-59259-508-2
eBook Packages: Springer Protocols