Two-Dimensional Gel Electrophoresis-Based Proteomic Analysis of Brain Synapses
Neuroproteomic technologies are crucial for a deeper understanding of molecular mechanisms underlying neuronal plasticity and pathologies of the central nervous system. For a comprehensive neuroproteomic analysis, high-resolution high-throughput techniques are indispensable. The most commonly used system for 2D gel electrophoresis is based on the combination of isoelectric focussing and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Nevertheless, the analysis of complex samples derived from biochemically prepared synaptic fractions raises some crucial challenges. Therefore, we describe the general 2D gel electrophoresis procedure, including sample preparation, gel casting, electrophoresis condition, and the detection of proteins on the gel with mass spectrometry compatible silver staining, sypro-ruby or colloidal Coomassie Brilliant Blue.
Key words2D gel electrophoresis Sample preparation Isoelectric focussing SDS-PAGE Protein staining Image analysis
This work was supported by bilateral programs of the Deutsche Forschungsgemeinschaft (DFG 444 CHL-113/32/0-1) and the Bundesministerium für Bildung und Forschung (BMBF CHL 06/027) with Conicyt (Chile), Proyecto Anillo 09-06 (PBCT, Conicyt, Chile) and the European Union (ZVOH-TP3/2, ZVOH 6/1).
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