title: Simultaneous Two-Dimensional Difference Gel Electrophoresis (2D-DIGE) Analysis of Two Distinct Proteomes.
abstract: This chapter describes the basics, applications, and limitations of two-dimensional gel electrophoresis (2DE) and two-dimensional difference gel electrophoresis (2D-DIGE) for multiplex analysis of distinct proteomes. We also propose a basic protocol for 2D-DIGE, technique that allows the analysis of paired protein extracts, which are labeled with fluorescent Cy3 and Cy5 dyes and electrophoresed with a Cy2-labeled standard extract on the same 2DE gels. Scanning the gels at wavelengths specific for each dye allows direct overlay the two different proteomes and the differences in abundance of specific protein spots can be determined.
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, Campinas, SP, 13083-862, Brazil.
authors.forename: Adriano
authors.initials: A
authors.lastname: Aquino
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, Campinas, SP, 13083-862, Brazil.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, Campinas, SP, 13083-862, Brazil. dmsouza@unicamp.br.|UNICAMP's Neurobiology Center, Campinas, Brazil. dmsouza@unicamp.br.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins-de-Souza
mesh_terms: D005456:Fluorescent Dyes; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012984:Software; D013223:Statistics as Topic; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D020543:Proteome
keywords: 2D difference gel electrophoresis; 2D-DIGE; 2DE; CyDyes; Proteomics