title: Protein profiling using two-dimensional difference gel electrophoresis (2-D DIGE).
abstract: 2-D DIGE relies on pre-electrophoretic labeling of samples with one of three spectrally distinct fluorescent dyes, followed by electrophoresis of all samples in one 2-D gel. The dye-labeled samples are then viewed individually by scanning the gel at different wavelengths, which circumvents problems with gel-to-gel variation and spot matching between gels. Image analysis programs are used to generate volume ratios for each spot, which essentially describe the intensity of a particular spot in each test sample, and thus enable protein abundance level changes to be identified and quantified. This unit describes the 2-D DIGE procedure including sample preparation from various cell types, labeling of proteins, and points to consider in the downstream processing of fluorescently labeled samples.
authors: Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.
authors: Renata
authors: R
authors: Feret
authors: Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.
authors: Kathryn S
authors: KS
authors: Lilley
mesh_terms: D005456:Fluorescent Dyes; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins
keywords: 2-D gel electrophoresis; DIGE; proteomics; quantitative