1 Fluorescence Two-dimensional Differential Gel Electrophoresis

To detect the proteome differences between two samples, the 2D-PAGE methodology uses the comparison of at least two gel images. But due to many reasons, no two gels images are superimposable directly. “Average gel ” from several gels has to be made before they are compared. In order to increase the accuracy and the reproducibility of 2D-PAGE, a new methodology Fluorescence Differential Gel Electrophoresis is used. Two protein samples are labeled with two cyanine dyes: cy3 and cy5 separately before they are mixed and run on one same 2D gel. By scanning the gel with two different lasers, two gel pictures from the same 2D picture can be made and they are perfectly superimposed. Compared to the conventional “one-sample-per-gel ” 2D-PAGE, Fluorescence Differential Gel Electrophoresis works much better in comparing two protein samples. Using the method described, the proteome changes of yeasts that are grown in medium containing 16 kinds of metal salts have been determined. The results are significant