title: A Two-Dimensional Difference Gel Electrophoresis (2D-DIGE) Protocol for Studies of Neural Precursor Cells.
abstract: This chapter describes the basics of two-dimensional difference gel electrophoresis (2D-DIGE) for multiplex analysis of up to distinct proteomes. The example given describes the analysis of undifferentiated and differentiated neural precursor cells labelled with fluorescent Cy3 and Cy5 dyes in comparison to a pooled standard labelled with Cy2. After labelling, the proteomes are mixed together and electrophoresed on the same 2D gels. Scanning the gels at wavelengths specific for each dye allows direct overlay of the two different proteomes and the differences in abundance of specific protein spots can be determined through comparison to the pooled standard.
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, SP, Brazil. paulcguest@yahoo.com.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
mesh_terms: D000818:Animals; D002458:Cell Fractionation; D002478:Cells, Cultured; D007202:Indicators and Reagents; D020547:Lateral Ventricles / Q000166:cytology; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000302:isolation & purification; D058953:Neural Stem Cells / Q000737:chemistry*; D018874:Spheroids, Cellular; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000295:instrumentation / Q000379:methods*
chemical_list: D007202:Indicators and Reagents; D009419:Nerve Tissue Proteins
keywords: 2D-DIGE; Brain; CyDyes; Differentiation; Neural precursor cells; Proteomics; Psychiatric disorders