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Features and applications of blue-native and clear-native electrophoresis

Journal

PROTEOMICS
Volume 8, Issue 19, Pages 3974-3990

Publisher

WILEY
DOI: 10.1002/pmic.200800017

Keywords

Activity stain; Blue-native electrophoresis; Clear-native electrophoresis; Protein-protein interaction; Supercomplex

Funding

  1. Deutsche Forschungsgemeinschaft
  2. Sonderforschungsbereich 472
  3. Project P11
  4. Cluster of Excellence Macromolecular Complexes [EXC 115]

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1-D native electrophoresis is used for the separation of individual proteins, protein complexes, and supercomplexes. Stable and labile protein-protein interactions can be identified depending on detergent and buffer conditions. 1-D native gels are immediately applicable for in-gel detection of fluorescent-labeled proteins and for in-gel catalytic activity assays. 1-D native gels and blots are used to determine native mass and oligomeric state of membrane proteins. Protein extracts from 1-D native gels are used for generation of antibodies, for proteomic work, and for advanced structural investigations. 2-D separation of subunits of protein complexes by SDS-PAGE is mostly used for immunological and proteomic studies. Following the discussion of these general features, specific applications of native electrophoresis techniques in various research fields are highlighted: immunological and receptor studies, biogenesis and assembly of membrane protein complexes, protein import into organelles, dynamics of proteasomes, proteome and subproteome investigations, the identification and quantification of mitochondrial alterations in apoptosis, carcinogenesis, and neurodegenerative disorders like Parkinson's disease, Alzheimer's disease, and the vast variety of mitochondrial encephalomyopathies.

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