Two-Dimensional Polyacrylamide Gel Electrophoresis Using Flat-Bed Isoelectric Focusing in the First Dimension
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 method 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 polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate (SDS-PAGE) in the second dimension. This technique is described in Chapter 11 and is reviewed in refs. 2, 3, 4.
KeywordsUrea Rubber Electrophoresis Syringe Polypeptide
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