(188bm) Role of Electrical Fields in the Pre-Treatment of Polyacrylamide Gels for Enhancing Protein Separations

Authors: 
Haris, A., Tennessee Technological University
Arce, P. E., Tennessee Technological University
Sanders, J. R., Tennessee Technological University
Protein electrophoresis is a commonly used technique to separate proteins based on their charge, molecular weight, mobility, etc. The technologies and methods associated with the electrophoresis technique are continuing to be developed to optimize the separation process and to improve the resulting resolution. Polyacrylamide gel electrophoresis (PAGE) is a widely used technique in protein separations. Different techniques have been developed for modifying the porous structure of polyacrylamide gel, such as varying the concentration of acrylamide, using a templating technique, and introducing nanoparticles to the gel, to improve the separation of different protein sizes. In this research, the effect of electrical fields in pretreating the gels, i.e. pre-electrophoresis, to enhance protein separation was investigated. Pre-electrophoresis is the application of an electrical field with constant voltage to the gel before protein separations by electrophoresis take place. The influence of pre-electrophoresis was studied by comparing the separation results of multiple proteins of different sizes in two types of gel samples with the same acrylamide concentration. The only difference was that one gel sample was subjected to pre-electrophoresis prior to protein separation, while the other was directly used for protein separation without the pre-electrophoresis step. Three different gel concentrations were tested: 6%, 9% and 12%. The results showed that the application of the pre-electrophoresis had a significant effect on the protein separations. The pre-electrophoresis technique enabled the resolution of all 10 proteins within a ladder of different sizes in polyacrylamide gels for low acrylamide concentration i.e. 6% and 9%, while without this technique only 12 % was able to separate all proteins. The novel effect of pre-electrophoresis has the great potential of reducing considerably the amount of gel material needed to conduct a separation.