Two-dimensional differential gel

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. 

Clii omy BA, Gonzales AD, Perkins J, Choi MW, Corzett MH, Chang BC, Corzett CH, McCutchen-Maloney SL (2004) Proteomic analysis of human serum by two-dimensional differential gel electi opho-resis after depletion of liigh-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. 

One way of linearizing the problem is to use the method of least squares in an iterative linear differential correction technique (McCalla, 1967). This approach has been used by Taylor et al. (1980) to solve the problem of modeling two-dimensional electrophoresis gel separations of protein mixtures. One may also treat the components—in the present case spectral lines—one at a time, approximating each by a linear least-squares fit. Once fitted, a component may be subtracted from the data, the next component fitted, and so forth. To refine the overall fit, individual components may be added separately back to the data, refitted, and again removed. This approach is the basis of the CLEAN algorithm that is employed to remove antenna-pattern sidelobes in radio-astronomy imagery (Hogbom, 1974) and is also the basis of a method that may be used to deal with other two-dimensional problems (Lutin et al., 1978 Jansson et al, 1983). 

Fig. 6. Two-dimensional polyacrylamide gel separation of hepatic proteins obtained from control and Entreated largemouth bass (injected IP with 2.5 mg/kg). The separation in the first dimension was on a pH 4-7 immobilized pH gradient strip and in the second dimension on an 8-16% gradient gel. A small segment of the gel is expanded for visual inspection. Arrows point to proteins that are differentially expressed in either E2 treated or control fish.

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. 

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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