Agarose-2D-DIGE: Fluorescent 2-D differential gel electrophoresis (2-D DIGE) with agarose gels in the first-dimensional isoelectric focusing (IEF)

Abstract

A two-dimensional gel electrophoresis (2-DE) method that uses an agarose isoelectric focusing (IEF) gel in the first dimension (agarose 2-DE) offers advantages over the more widely used immobilized pH gradient 2-DE for separating high-molecular-mass proteins (100-500kDa) and for having a higher loading capacity (1.5mg in total). We have applied the Fluorescent 2-D differential gel electrophoresis (2-D DIGE) to our agarose 2-DE system. This allowed us to see clear differences in the 2-DE patterns from liver extracts of a diabetic model Otsuka Long-Evans Tokushima Fatty (OLETF) and its control Long-Evans Tokushima Otsuka (LETO) rats including changes in the amounts of several proteins larger than 100kDa. The combined method would increase its power to detect changes in disease proteomics.