Gel Electrophoresis, 2D-difference

Abstract

Fluorescence two-dimensional difference gel electrophoresis (2D-DIGE) is a recently developed 2D-gel-based proteomics technique that can be applied to provide a sensitive, rapid, and quantitative analysis of differential protein expression between two or more biological samples. This technique utilizes chemical derivatives of spectrally distinct fluorescent cyanine dyes (Cy2, Cy3, and Cy5) to covalently label different samples prior to mixing and separation on the same 2-DE gels. This differential labeling and mixing of samples helps avoid the gel-to-gel variation inherent in conventional 2-DE-based expression profiling and reduces the number of gels that need to be run. Since the technique is also compatible with downstream identification of protein spots from gels by mass spectroscopy, it is particularly useful for reproducible, high-throughput proteomics.