Three-Dimensional Electrophoresis for Quantitative Profiling of Complex Proteomes


  • Mauro, S. , Colignon, B. , Dieu, M. , Delaive, E. , Dieu, M. & Raes, M. (2015). Three-Dimensional Electrophoresis for Quantitative Profiling of Complex Proteomes In: Proteomic Profiling: Methods and Protocols, Methods in Molecular Biology, Anton Posch. New York, Springer Science+Business Media New York 2015, 1295,
Type Book Section
Year 2015
Chapter title Three-Dimensional Electrophoresis for Quantitative Profiling of Complex Proteomes
Book title Proteomic Profiling: Methods and Protocols, Methods in Molecular Biology
Editor Anton Posch
City New York
Publisher Springer Science+Business Media New York 2015
Volume 1295
Abstract Quantitative 2D-gel-dependent proteomics became feasible with 2D fluorescence difference gel electrophoresis (2D-DIGE), and this technique has gained wide acceptance because it has eliminated the gel to gel variations and greatly facilitated the quantitative comparisons across gels for many different experimental conditions. However, the co-migration of several proteins in the same spot is still a major limitation which detracts from the accuracy of comparative quantification and prevents unambiguous post-translational modifications (PTMs) detection. A protocol based on traditional polyacrylamide gel IEF sample fractionation, and followed by two consecutive SDS-PAGE electrophoreses alleviates co-migration limitations. The use of two different buffer systems for SDS-PAGE is central to the proposed approach.
Fichier
Lien DOI 10.1007/978-1-4939-2550-6_30
Authors Mauro, S. , Colignon, B. , Dieu, M. , Delaive, E. , Dieu, M. & Raes, M.