Quantitative gel electrophoresis

Gel electrophoresis is the most widely used technique for the analysis of protein samples, and there are a variety of methods that can be used to investigate single proteins as well as highly complex protein mixtures. To ensure reproducible and reliable separations of such samples and the resolution of distinct protein species, there has been a substantial amount of research dedicated to optimising methods to the refined techniques available today. There are thus a number of factors that have a marked influence on the practice of quantitative proteomics. The procedures and reagents involved in preparing a protein sample can have a significant effect on the composition of the proteome and/or its resolution by electrophoresis. Furthermore, since most proteins are colourless, a protein stain is required to detect the resolved proteome; thus, it is essential that the characteristics of the stain enable optimal detection regardless of protein type and/or concentration. Notably, to obtain reliable quantitative data, the approach by which images are acquired is equally important.