Difference gel electrophoresis

Unlu, M., Morgan, M.E., Minden, J.S. (1997). Difference gel electrophoresis a single gel method for detecting changes in protein extracts. Electrophoresis 18, 2071-2077. 

Kondo T, Hirohashi S. (2006) Application of highly sensitive fluorescent dyes (CyDye DICE Fluor saturation dyes) to laser micro-dissection and two-dimensional difference gel electrophoresis (2D-DICE) for cancer proteomics. NatProtoc 1,2940-56. 

Shaw J, Rowhnson R, Nickson J, et al. (2003) Evaluation of saturation labelling two-dimensional difference gel electrophoresis fluorescent dyes. Proteomics 3, 1181-95. 

Sitek B, Luttges J, Marcus K, et al. (2005) Application of fluorescence difference gel electrophoresis saturation labelling for the analysis of microdissected precursor lesions of pancreatic ductal adenocarcinoma. Proteomics 5, 2665-79. 

Fujii K, Kondo T, Yokoo H et al. Proteomic study of human hepatocellular carcinoma using two-dimensional difference gel electrophoresis with saturation cysteine dye. Proteomics 2005 5 1411-1422. 

DE, two-dimensional gel electrophoresis WB, Western blot MS, mass spectrometry LC, liquid chromatography 2DE DICE, two-dimensional difference gel electrophoresis, SRM, selected-reaction monitoring MRM, multiple reaction monitoring AQUA, absolute protein quantitation SMIM, selected MS/MS ion monitoring. ... 

At present, there are advanced difference gel electrophoresis (DOGE) Systems and 2-D fluorescence difference gel electrophoresis (2-D DIGE) which enable the analyst to use simultaneously modern (more precise) methods of fluorescent analysis with 2-D electrophoresis (using internal patterns), aided by a fully integrated bioinformatics system. Such systems allow more complete differential protein analysis, while the application of internal standards eliminates differentiation between the intervals, thus ensuring that even the smallest differences will be detected irrespective of the multitude of components. This guarantees reproducibility of results and their statistical reliability. Such assays are one of the platforms employed in the research based on the proteomics method. 

Hobson, D.J., Rupa, P., Diaz, G.J., Zhang, H., Yang, M., Mine, Y., Turner, P.V., Kirby, G.M. 2007. Proteomic analysis of ovomucoid hypersensitivity in mice by two-dimensional difference gel electrophoresis (2D-DIGE). Food Chem Toxicol 45 2372-2380. 

Liang CR, Leow CK, Neo JC, Tan GS, Lo SL, Lim JW, et al. Proteome analysis of human hepatocellular carcinoma tissues by two-dimensional difference gel electrophoresis and mass spectrometry. Proteomics 2005 5(8) 2258-2271. 

D difference gel electrophoresis (DIGE), etc. To learn the handling of this software, several ProteinScape training courses took place at periodic intervals and more will be held in the future. 

In the last years the application of two-dimensional electrophoresis (2-DE) has often been declared outdated and a new century of gel-free proteomics was announced. Nevertheless, 2-DE is still the method of choice when analyzing complex protein mixtures. With a separation of10 000 proteins, 2-DE gives access to high-resolution proteome analysis. Continuous development has consolidated 2-DE application in proteomics, where the introduction of difference gel electrophoresis (DICE) is the latest improvement. DICE is based on fluorescently tagging all proteins in each sample with one set of matched fluorescent dyes designed to minimally interfere with protein mobility during 2-DE. 

Figure 3.2 Difference gel electrophoresis (DIGE). Ettan DIGE workflow three-color and two-color experiments including the internal standard. For fluorescence proteins tagging, two different CyDyes techniques are available. Minimal fluors allow consideration of three different CyDyes (Cy2, Cy3 and Cy5) in a multiplexing experiment.

Difference Gel Electrophoresis Next Generation of Protein Detection in 2-DE 45... 

Alban, A., David, S. O., Bjorkesten, L, Andeesson, C., Sloge, E., Lewis, S., Currie, I. (2003). A novel experimental design for comparative two-dimensional gel analysis two-dimensional difference gel electrophoresis incorporating a pooled internal standard. Proteomics 3, 36-44. 

Determining a significant change in protein expression with DeCyder during a pairwise comparison using two-dimensional difference gel electrophoresis. Proteomics 4, 1421-1432. 

Peeieeer, K., Meyer, H. E., Eggert, A., Schramm, A. (2005a). Identification of dynamic proteome changes upon ligand activation of Trk-receptors using two-dimensional fluorescence difference gel electrophoresis and mass spectrometry. Mol. Cell Proteomics 4, 291-299. 

Despite its lower resolution compared to 2-DE, 2-DB can also be applied in combination with the difference gel electrophoresis technique (DIGE) (Urdu et al. 1997 Reinders et al. 2006b), enabling the highly reproducible differential analysis of biological membrane samples. 

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