Two-dimensional gel electrophoresis-based proteomic analysis of brain synapses

Karl Heinz Smalla, Úrsula Wyneken

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

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.

Original languageEnglish
Title of host publicationNeuroproteomics
EditorsKa Wan Li
Pages95-113
Number of pages19
DOIs
StatePublished - 2011
Externally publishedYes

Publication series

NameNeuromethods
Volume57
ISSN (Print)0893-2336
ISSN (Electronic)1940-6045

Keywords

  • 2D gel electrophoresis
  • Image analysis
  • Isoelectric focussing
  • Protein staining
  • Sample preparation
  • SDS-PAGE

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