Capabilities using 2-D DIGE in proteomics research : the new gold standard for 2-D gel electrophoresis.
The use of two-dimensional gel electrophoresis for differential analysis in proteomics was revolutionized by the introduction of 2-D fluorescence difference gel electrophoresis (2-D DIGE). This fluorescence-based technique allows the use of multiplexed samples and an internal standard that virtually eliminates gel-to-gel variability, resulting in increased confidence that differences found between samples are due to real induced changes, rather than inherent biological variation or experimental variability. 2-D DIGE has quickly become the "gold standard" for gel-based proteomics for studying tissues, as well as cell culture and bodily fluids such as serum or plasma. This chapter will describe the basic 2-D DIGE technique using minimal labeling, image acquisition using high-quality fluorescence scanners, and software capable of analyzing the multiplexed images and normalizing the data using the internal standard.
Rozanas
Christine R
CR
GE Healthcare, 800 Centennial Ave., Piscataway, NJ, USA.
Loyland
Stacey M
SM
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents; D007525:Isoelectric Focusing; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D012984:Software
D007202:Indicators and Reagents; D011506:Proteins