title: Two-dimensional difference gel electrophoresis.
abstract: Two-dimensional difference gel electrophoresis (2D DIGE) is a modified form of 2D electrophoresis (2DE) that allows one to compare two or three protein samples simultaneously on the same gel. The proteins in each sample are covalently tagged with different color fluorescent dyes that are designed to have no effect on the relative migration of proteins during electrophoresis. Proteins that are common to the samples appear as 'spots' with a fixed ratio of fluorescent signals, whereas proteins that differ between the samples have different fluorescence ratios. With the appropriate imaging system, DIGE is capable of reliably detecting as little as 0.5 fmol of protein, and protein differences down to +/- 15%, over a >10,000-fold protein concentration range. DIGE combined with digital image analysis therefore greatly improves the statistical assessment of proteome variation. Here we describe a protocol for conducting DIGE experiments, which takes 2-3 d to complete.
authors.affiliation: Department of Biological Science, Carnegie Mellon University, Mellon Institute, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: Surya
authors.initials: S
authors.lastname: Viswanathan
authors.forename: Mustafa
authors.initials: M
authors.lastname: Unlü
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Minden
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005456:Fluorescent Dyes; D011506:Proteins / Q000032:analysis*
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins