ABSTRACT
Two-dimensional gel electrophoresis (2DE) was developed in the late-1960s as a method for enhanced protein separation. Since that time the method has rapidly evolved with ever-increasing protein separation capabilities and protocols for protein detection. In 2DE, proteins are separated in two dimensions based on the intrinsic properties of each protein, which are then stained for visualization and quantication (Figure 20.1). By comparing the intensities of the stained protein spots, their relative amounts are estimated. Protein spots of interest that display differential levels or modied migration patterns in a biological comparison are excised from the gel, digested into peptides, and analyzed by MS to obtain protein identication.
