Diffusing effect of DTT and 2-ME during SDS-PAGE electrophoresis / 医学研究生学报

ABSTRACT

Objective: Dithiothreitol(DTT) and ?-Mercaptoethanol(2-ME) are important reducing agents for SDS-PAGE.This study is to observe the diffusing effect of DTT and 2-ME during electrophoresis,and to find a way of avoiding this effect.Methods: We placed protein samples containing reducing agents and non-reduced protein samples separately in the sample wells at intervals for SDS-PAGE electrophoresis,and determined whether or not the electrophoretic lanes of the non-reduced samples were interfered by the adjacent lanes.Results: DTT and 2-ME diffused to the neighboring lane,so that the non-reduced samples were reduced partially.The spreading effect was positively correlated with the content of the reducing agent.Conclusion: DTT and 2-ME have a diffusing effect during SDS-PAGE electrophoresis.In separating the reduced and non-reduced proteins in the same gel at the same time,at least a blank lane should be set up in between them in order to avoid the diffusing effect.