Two-dimensional differential gel electrophoresis

Tonge R, Shaw J, Middleton B et al. Validation and development of fluorescence two-dimensional differential gel electrophoresis proteomics technology. Proteomics 2001 1 377-396. 

ZvELEBiL, M.)., Cramer, R., Waterfield, M. D., Timms, ). E. (2002). Evaluation of two-dimensional differential gel electrophoresis for proteomic expression analysis of a model breast cancer cell system. Mol. Cell Proteomics 1, 91-98. 

Rowlinson, R., Rayner, S., Young, )., PoGNAN, F., Hawkins, E., Currie, I., Davison, M. (2001). Validation and development of fluorescence two-dimensional differential gel electrophoresis proteomics technology. Proteomics 1, 377-396. 

Chromy BA, Gonzales AD, Perkins J, Choi MW, Corzett MH, Chang BC, Corzett CH, McCutchen-Maloney SL (2004) Proteomic analysis of human semm by two-dimensional differential gel electrophoresis after depletion of high-abundant proteins. J Proteome Res 3 1120-1127. 

Alfonso, P., Nunez, A., Madoz-Gurpide, J., Lonbardia, L., Sanchez, L and Casal, J. I. (2005) Proteomic expression analysis of colorectal cancer by two-dimensional differential gel electrophoresis. Proteomics 5, 2602-11. 

CP from all species analyzed so far occurs as multiple basic isoforms with isoelectric points around 9 (Takahashi et al, 1988a). Using two-dimensional (2D) gel electrophoresis it has been established that CP is expressed in up to three major isoelectric variants (a, P, 7 see Fig. 2) of similar molecular mass, ranging from pi s of 9.9 to 9.4 (Gimona et al, 1992). The isoforms appear progressively with the differentiation in chicken gizzard, porcine stomach (Gimona et al,... 

Two dimensional polyacrylamide gel electrophoresis of helminth and mammalian tubulin revealed a difference in their a-subunits. This was confirmed by limited proteolytic peptide mapping where it was consistently observed that some peptides were novel to each tubulin.It was suggested that the difference in the cytoplasmic a-tubulin subunit described compared with that of mammalian cells could account for the selective toxicity of the benzimidazole carbamates without the need to postulate differential pharmacokinetics between parasite and host. 

Van den Bergh, G., Clerens, S., Vandesande, F., and Arckens, L. (2003) Reversed-phase high-performance liquid chromatography prefractionation prior to two-dimensional difference gel electrophoresis and mass spectrometry identifies new differentially expressed proteins between striate cortex of kitten and adult cat. Electrophoresis 24, 1471-81. 

For reproducible expression analysis and protein quantification MS methods based on isotopic labeling are available. They were designed in conjunction with two or more dimensional chromatographic peptide separation coupled online to MS and require advanced bioinformatics input to analyze the complex data sets in a reasonable time frame. This is also true for the alternative fluorescence-based technology of differential gel electrophoresis (DIGE Fig. 10.6) with tailor-made software which allows statistical validation of multiple data sets. 

Quantification by Two-Dimensional Differential Imaging Gel Electrophoresis 2D-DIGE is a more precise version of the 2-DE approach. This method uses multicolored cyanine dyes to label proteins in the two experimental samples... 

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