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.
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We would like to thank Drs. Aude Foucher and Jolyne Drummelsmith for previous optimization with our FFE. MO is member of the CIHR Group on Host–Pathogen Interactions and of the Centre for Host–Parasite Interactions “Programme Regroupements Statégiques” of the Fonds du Québec pour la Recherche sur la Nature et les Technologies. This work was funded by a CIHR grant to MO. MO holds the Canada Research Chair in Antimicrobial Resistance.
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