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Effects of different current densities on efficiency of electrotransfer for the large-molecular-weight protein in Western blot assay

Xie Lan1,Ai Hua2 (1.Nutrition and Biochemistry Section,Institute of Sports Medicine,Third Hospital, Peking University,Beijing 100191,China;2.Nutrition and Exercise Section,Obesity and Metabolic Diseases Research Centre,Peking University,Beijing 100191,China)  
Objective To study the effects of different current densities on electrotransfer efficiency for the large-molecular-weight protein in Western blot assay.Methods (36-250)×103 protein markers were migrated on 8% SDS-PAGE gel and then wetly electrotransferred at constant current density of 100,200,300,400,500 mA,respectively,for 2 hours.Results The 250×103 marker was not totally transferred at 100 and 200 mA with visual remains on the gel.However,it was mostly or fully transferred to the membrane at 300-500 mA without any visual remains on the gel.Conclusion The efficiency of electroblotting of a large-molecular-weight protein might be affected by the current density and 300-400 mA could be optimal constant current densities for the wet electroblotting of the proteins as large as 250×103 from 8% SDS-PAGE gel.
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