上槽缓冲容量不足导致双向电泳纵向拖尾原因初探

Insufficient buffer capacity in up chamber leads to vertical streaking of two-dimensional electrophoresis

目的探讨上槽缓冲容量对双向电泳纵向拖尾的影响。方法保持上槽缓冲液体积不变,分别采用1×buffer电泳2块或1块SDS胶,或1.5×、2×、3×buffer电泳2块胶,比较纵向拖尾情况。结果 1×buffer电泳2块胶,整块胶出现严重纵向拖尾,电泳1块胶时无纵向拖尾。1.5×buffer电泳2块胶,纵向拖尾只存在于高分子量区,中低分子量区蛋白点呈圆形,无拖尾。2×buffer电泳2块胶,高分子量区纵向拖尾较1.5×buffer进一步减少,中低分子量区圆形蛋白点增多。3×buffer电泳2块胶时,纵向拖尾完全消除。结论上槽缓冲液缓冲容量不足导致双向电泳纵向拖尾。

Objective To study the effect of buffer capacity in up chamber on vertical streaking（VSTK） of two-dimensional electrophoresis.Methods The same buffer volume was kept in up chamber,VSTK was compared by running two SDS gels or single one in 1× Laemmli buffer（buffer）,or two ones in 1.5×,2× and 3× buffer in up chamber respectively.Results VSTK appeared severely when two gels were run in 1× buffer,while disappeared when one did.When two gels were run in 1.5×buffer,VSTK only existed in the region of high molecular weight,and protein spots in the region of medium and low molecular weight were round without streaking.When two gels were run in 2× buffer,compared with 1.5× buffer,the VSTK in the region of high molecular weight was further improved,and the round protein spots in the region of medium and low molecular weight increased.VSTK disappeared completely when two gels were run in 3×buffer.Conclusion Insufficient buffer capacity in up chamber may lead to vertical streaking.