Gel Electrophoresis of Proteins: High‐resolution Two‐dimensional

Abstract

The technique of high‐resolution two‐dimensional gel electrophoresis separates proteins according to both isoelectric point and molecular weight, giving it unsurpassed resolving power.

Keywords: 2D PAGE; isoelectric focusing (IEF); nonequilibrium pH gradient; immobilized pH gradient

Figure 1.

Noncultured normal human keratinocyte proteins separated using (a) IPG (pH 3–10) and (b) carrier ampholytes (pH 3.5–10). A few proteins are indicated for references. Part (a) is reproduced from Bjellqvist et al. .

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References

Bjellqvist B, Ek K, Righetti PG et al. (1982) Isoelectric focusing in immobilized pH gradients: principle, methodology and some applications. Journal of Biochemical and Biophysical Methods 6: 317–339.

Bjellqvist B, Basse B, Olsen E and Celis JE (1994) Reference points for comparisons of two‐dimensional maps of proteins from different human cell types defined in a pH scale where isoelectric points correlate with polypeptide compositions. Electrophoresis 15: 529–539.

Celis JE, Ratz G, Basse B et al. (1997) High‐resolution two‐dimensional gel electrophoresis of proteins: isoelectric focusing (IEF) and nonequilibrium pH gradient electrophoresis (NEPHGE). In: Celis JE, Carter N, Hunter T et al. (eds) Cell Biology, A Laboratory Handbook, vol. 4, pp. 375–385. New York: Academic Press.

Celis JE, Østergaard M, Jensen NA et al. (1998) Human and mouse proteomic databases: novel resources in the protein universe. FEBS Letters 430: 64–72.

Görg A and Weiss W (1997) High‐resolution two‐dimensional electrophoresis of proteins using immobilised pH gradient. In: Celis JE, Carter N, Hunter T et al. (eds) Cell Biology, A Laboratory Handbook, vol. 4, pp. 386–397. New York: Academic Press.

Klose J (1975) Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals. Humangenetik 26: 231–243.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

O’Farrell PH (1975) High resolution two‐dimensional electrophoresis of proteins. Journal of Biological Chemistry 250: 4007–4021.

O’Farrell PZ, Goodman HM and O’Farrell PH (1977) High resolution two‐dimensional electrophoresis of basic as well as acidic proteins. Cell 12: 1133–1141.

Further Reading

Celis JE and Bravo R (1984) Two‐dimensional Gel Electrophoresis of Proteins: Methods and Applications. New York: Academic Press.

Link AJ (ed.) (1998) 2‐D Proteome Analysis Protocols. Methods in Molecular Biology. Totowa, NJ: Humana Pess.

Wilkins MR, Williams KL, Appel RD and Hochstrasser DF (eds) (1997) Proteomic Research: New Frontiers in Functional Genomics (Principles and Practice). Berlin: Springer‐Verlag.

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How to Cite close
Celis, Julio E, and Gromov, Pavel S(Apr 2001) Gel Electrophoresis of Proteins: High‐resolution Two‐dimensional. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002684]