Two-Dimensional Gel Electrophoresis-Based Proteomic Analysis of Brain Synapses

  • Karl-Heinz SmallaEmail author
  • Ursula Wyneken
Part of the Neuromethods book series (NM, volume 57)


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 words

2D 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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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Leibniz Institute for NeurobiologyMagdeburgGermany

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