Abstract
Two-dimensional electrophoresis (2-DE) is the leading tool in proteomics research today, capable of visualising many components of complex proteomes in a single gel (Gorg et al. 2000). This tool separates proteins by pI (isoelectric point) and molecular weight producing a pattern of spots on an SDS-PAGE (polyacrylamide gel electrophoresis) gel, which can be visualised via a range of staining and labelling systems. Protein spot patterns can be compared between different samples of interest, e.g. normal versus diseased, to determine which protein spots demonstrate changes in abundance. The most common 2-DE work system incorporates the analysis/identification of differentially expressed proteins of interest using Mass Spectrometry. Figure 15.1 illustrates a 2-DE workflow from sample preparation to Mass Spectrometry analysis.
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Prime, J., Alban, A., Hawkins, E., Hughes, B. (2004). The Quantitative Advantages of an Internal Standard in Multiplexing 2D Electrophoresis. In: Kamp, R.M., Calvete, J.J., Choli-Papadopoulou, T. (eds) Methods in Proteome and Protein Analysis. Principles and Practice. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-08722-0_15
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DOI: https://doi.org/10.1007/978-3-662-08722-0_15
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