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State-of-the-art two-dimensional gel electrophoresis: a key tool of proteomics research

Published in Nature Protocols. 2006, vol. 1, no. 2, p. 812-23
Abstract Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is the most popular and versatile method of protein separation among a rapidly growing array of proteomics technologies. Based on two distinct procedures, it combines isoelectric focusing (IEF), which separates proteins according to their isoelectric point (pI), and SDS-PAGE, which separates them further according to their molecular mass. At present, 2D-PAGE is capable of simultaneously detecting and quantifying up to several thousand protein spots in the same gel image. Here we provide comprehensive step-by-step instructions for the application of a standardized 2D-PAGE protocol to a sample of human plasma or cerebrospinal fluid (CSF). The method can be easily adapted to any type of sample. This four-day protocol provides detailed information on how to apply complex biological fluids to an immobilized dry strip gel, cast home-made gradient acrylamide gels, run the gels, and perform standard staining methods. A troubleshooting guide is also included.
Keywords Blood Proteins/chemistryCerebrospinal Fluid/chemistryElectrophoresis, Gel, Two-Dimensional/methodsHumansProteomics/methods
PMID: 17406312
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Research groups Chimie et protéomique clinique (270)
Groupe de Protéomique biomédicale (635)
Groupe Pierre Burkhard (neurologie) (154)
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CARRETTE, Odile et al. State-of-the-art two-dimensional gel electrophoresis: a key tool of proteomics research. In: Nature Protocols, 2006, vol. 1, n° 2, p. 812-23. https://archive-ouverte.unige.ch/unige:33754

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Deposited on : 2014-01-28

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