Two-dimensional polyacrylamide gel electrophoresis 2D-PAGE

Two-Dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE lEF followed by SDS-PAGE)... 

Two Dimensional-Polyacrylamide Gel Electrophoresis (2D-PAGE) is the most powerful tool to separate the different components of complex protein mixtures. Proteins are first separated according to their p.I. in IEF (first dimension), then a further orthogonal separation according to the MW is performed by SDS-PAGE (second dimension). In this way all the protein components of a biological sample can be virtually separated. 

In proteomics the sample complexity is often high and requires high-resolution analytical separation techniques. Two-dimensional polyacrylamide gel electrophoresis (2D PAGE) fulfills this requirement and has been established as a core technique for complex protein mixtures. However, the small dynamic range and the inability to resolve very small and large proteins and proteins with extreme pi values have been serious limitations. 

The first step in the physiological proteomics analysis of S. aureus is the establishment of a comprehensive two-dimensional (2D) gel proteome map that should cover most of the cytoplasmic proteins expressed under specific circumstances see Fig. 1). For physiological studies, the highly sensitive two-dimensional polyacrylamide gel electrophoresis (2D PAGE) originally invented by O Farrel (2) and Klose (3) is still the state of the art. With our standard 2D gel system (pH 4-7), we identified 473 proteins that cover about 40% of the cytoplasmic proteome of S. aureus predicted for this proteomic window (4). The proteins are associated with various cellular functions ranging from the transcriptional and translational machinery, citrate cycle, glycolysis, and fermentation pathways to biosynthetic pathways of nucleotides, fatty acids, and cell wall components (4,5). 

Two-dimensional, polyacrylamide gel electrophoresis (2D PAGE) is an open assay approach, which permits the separation and detection of proteins from a wide variety of sources without the need for any prior... 

Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), which has been successfully used to establish a correlation between adsorbed proteins and in vivo behavior of polystyrene carriers coated with Poloxamers (Blunk etal, 1994) could be a helpful tool to study competitive plasma protein adsorption on PEG-coated nanospheres as a function of PEG chain length and surface density. Indeed, the relevance of the adsorbed plasma proteins (and their ratios and conformations) with respect to the in vivo fate of the particles remains to be uncovered. 

Two-dimensional polyacrylamide gel electrophoresis (2D PAGE) is a powerful approach to the analysis of complex protein mixtures as polypeptides are separated on the basis of two electrophoretic properties ... 

---