Fluorescent difference gel electrophoresis.

INTRODUCTION A bottleneck for high-throughput proteomic studies is image analysis. In conventional two-dimensional (2D) methodology, protein samples are separated on individual gels, stained, and quantified, followed by image comparison with computer-aided image analysis programs. Since different images are not always perfectly superimposable, image analysis is very time consuming. Fluorescent difference gel electrophoresis shortens this procedure (Ünlü et al. 1997) by independently labeling samples in vitro before IEF using two different fluorescent cyanine dyes, then mixing and separating the samples on a single 2D gel. Subsequent image analysis enables the differences (e.g., upor down-regulated proteins) between the samples to be visualized.