Two-Dimensional Polyacrylamide Gel Electrophoresis – A Practical Perspective

Twodimensional electrophoresis, abbreviated as 2-DE, is one of the most powerful and common tools for separation and fractionation of complex protein mixture extracted from tissues, cells, and other biological specimens. It is an orthogonal technique that allows separation of thousands of proteins in one gel and in a two tandem electrophoretic steps where a major proportion of proteins can be resolved for further analysis. Since the pioneer development and modification of high resolution 2-DE by (O'Farrell, 1975), the usage of this technique is increasing and proven to be effective. In fact, the modification set by O’Farrell empowered this technique to resolve up to 5000 spots in a uniform distribution and on a single gel. Protein spot can be seen as little as one disintegration per min by autoradiology or recent quantitative dye. This high reproducibility of separation allows comparative matching for detection of dysregulated proteins.