2-D Electrophoresis Using Carrier Ampholytes in the First Dimension (IEF)
Two-dimensional electrophoresis (2-DE) is potentially the most powerful technique known for resolving complex protein mixtures. Proteins can be purified to homogeneity in one step in most cases. The applications for 2-DE are numerous and diverse. They include the analysis of protein patterns in various organisms and tissues, monitoring of the purity of fractions derived from chro-matography (or other separation methods), and the determination of the characteristics of a particular protein, such as isoelectric point, molecular weight of subunits, and number of isoforms.
KeywordsCasting System Carrier Ampholyte Complex Protein Mixture Anode Solution Acrylamide Solution
- 1.Aebersold, R. H. (1989) Internal amino acid sequence analysis of proteins after in situ protease digestion on nitrocellulose, in A Practical Guide to Protein and Peptide Purification for Microsequencing (Matsudaira, P., ed.), Academic, San Diego, CA, pp. 73–81.Google Scholar
- 7.Dunbar, B. S. (1987) Basic theories and principles of electrophoresis, in Two-Dimensional Electrophoresis and Immunological Techniques. Plenum, NY, pp. 1–21.Google Scholar
- 8.Blomberg, A., Blomberg, L., Norbeck, J., Fey, S. J., Mose Larsen, P., Larsen, M., Roepstorff, M., Degand, H., Boutry, M., Posch, A., and Gorg, A. (1995) Interlaboratory reproducibility of yeast protein patterns analyzed by immobilized pH gradient two-dimensional gel electrophoresis. Electrophoresis 16, 1935–1945.Google Scholar