Separation of basic proteins from Leishmania using a combination of Free flow electrophoresis (FFE) and 2D electrophoresis (2-DE) under basic conditions

Methods Mol Biol. 2015:1201:247-59. doi: 10.1007/978-1-4939-1438-8_15.

Abstract

Basic proteins, an important class of proteins in intracellular organisms such as Leishmania, are usually underrepresented on 2D gels. This chapter describes a method combining basic proteins fractionation using Free flow electrophoresis in isoelectric focusing mode (IEF-FFE) followed by protein separation using two-dimensional gel electrophoresis (2-DE) in basic conditions. The combination of these two techniques represents a great improvement for the visualization of Leishmania proteins with basic pI using 2D gels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Buffers
  • Calibration
  • Electrophoresis / instrumentation
  • Electrophoresis / methods*
  • Electrophoresis, Gel, Two-Dimensional / instrumentation
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Equipment Design
  • Image Processing, Computer-Assisted / methods
  • Isoelectric Focusing / methods
  • Leishmania* / growth & development
  • Protozoan Proteins / analysis
  • Protozoan Proteins / isolation & purification*

Substances

  • Buffers
  • Protozoan Proteins