Stopped Lower Buffer Flow Electrofractionation: Simple Electrofractionation for Complex Protein Mixture

Abstract

The standard electrofractionation technique simulated the liquid chromatography to create a continuous elution system. Though this continuous lower buffer flow electrofractionation system served as an excellent model for preparative electrophoresis, the restricted current flow caused by electric resistance arising from the column holder prevented the application to the purification of complex protein mixtures. To overcome the electric current problem, the stopped lower buffer flow electrofractionation (SFE) system is designed to work without using a column holder and use prestained markers as the detection tool of protein position during the electrofractionation process. Through this improvement, the SFE system widens the availability of electrofractionation to the purification of complex and low-concentration protein mixtures. In this report, the efficiency of this technique is shown through the partial purification of a 32P-labeled protein in the rat liver microsome.