Separation of basic proteins from Leishmania using a combination of Free flow electrophoresis (FFE) and 2D electrophoresis (2-DE) under basic conditions.
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.
Centre de Recherche en Infectiologie, CHU-Québec research centre, 2705 boul Laurier, Québec, Canada, G1V 4G2.
Marie-Christine
MC
Brotherton
Gina
G
Racine
Marc
M
Ouellette
D002021:Buffers; D002138:Calibration; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007525:Isoelectric Focusing / Q000379:methods; D007891:Leishmania / Q000254:growth & development; D015800:Protozoan Proteins / Q000032:analysis / Q000302:isolation & purification*
D002021:Buffers; D015800:Protozoan Proteins