Two-dimensional blue native/SDS gel electrophoresis of multiprotein complexes

Methods Mol Biol. 2012;869:317-37. doi: 10.1007/978-1-61779-821-4_27.

Abstract

The two-dimensional blue native/sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D BN/SDS-PAGE) is a method of choice for the investigation of protein complexes. This highly resolvent separation method is unique in that it facilitates the identification of many protein complexes simultaneously. Because of its simplicity and suitability, the 2D BN/SDS-PAGE can be now applied to a wide range of organisms, including bacteria, viruses, yeasts, animals, and plants. Moreover, recent modifications have made it possible to apply this method to the study of whole protein complexes of an organism. Here, we propose protocols for the investigation of the whole complexome of bacteria as well as eukaryotic cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Buffers
  • Caco-2 Cells
  • Cell Extracts / chemistry
  • Cell Extracts / isolation & purification
  • Coloring Agents / chemistry
  • Electrophoresis, Gel, Two-Dimensional / methods
  • Escherichia coli / chemistry
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / isolation & purification
  • Fungal Proteins / chemistry
  • Fungal Proteins / isolation & purification
  • Helicobacter pylori / chemistry
  • Humans
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / isolation & purification*
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification
  • Protein Structure, Quaternary
  • Proteolysis
  • Proteome / chemistry
  • Proteome / isolation & purification
  • Rosaniline Dyes / chemistry
  • Silver Nitrate / chemistry
  • Silver Staining
  • Trypsin / chemistry
  • Yarrowia / chemistry

Substances

  • Buffers
  • Cell Extracts
  • Coloring Agents
  • Escherichia coli Proteins
  • Fungal Proteins
  • Multiprotein Complexes
  • Peptide Fragments
  • Proteome
  • Rosaniline Dyes
  • Silver Nitrate
  • Trypsin
  • coomassie Brilliant Blue