【摘要】 目的研究Western blot方法中不同电流强度对较大分子蛋白质转膜效率的影响。方法利用(36～250)×103标志蛋白质,以8%SDS-PAGE凝胶分离,然后分别以100、200、300、400、500mA恒流湿法电转2h。结果 100、200mA恒流2h不能使250×103较大分子蛋白质全部转膜,凝胶有残留。300mA以上则可使大部分或全部转膜,胶上未见残留。结论电流强度对较大蛋白质转膜有影响。300～400mA可作为250×103左右较大分子蛋白质从8%SDS-PAGE凝胶转膜的最佳恒流强度。更多还原

【Abstract】 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.更多还原