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  • Title: Two-dimensional polyacrylamide gel electrophoresis using flat-bed isoelectric focusing in the first dimension.
    Author: Dunn MJ, Patel K.
    Journal: Methods Mol Biol; 1988; 3():217-31. PubMed ID: 21400166.
    Abstract:
    Electrophoretic techniques are, perhaps, the most important group of procedures for the separation and analysis of complex protein mixtures because of their high resolution capacity. This resolving power is maximized in high-resolution, two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) based on the technique developed by O'Farrell (1). This methodology involves a combination of a first-dimension separation in cylindrical isoelectric focusing (IEF) gels containing 9M urea and 2% (w/v) of the detergent NP-40 with discontinuous gradient polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) in the second dimension. Detailed reviews of the theoretical basis, methodological variations, and applications of this technology can be found in refs. 2-5.
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