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膜蛋白的苯酚-乙酸-尿素聚丙烯酰胺凝胶电泳

Phenol-acetic acid-urea polyacrylamide gel electrophoresis of membrane proteins.

作者信息

Gallagher S R, Leonard R T

出版信息

Anal Biochem. 1987 May 1;162(2):350-7. doi: 10.1016/0003-2697(87)90403-9.

Abstract

A phenol-acetic acid-urea polyacrylamide gel electrophoretic system (PAU-PAGE) was simplified by adaptation to a slab gel format, allowing the simultaneous comparison of up to 12 samples. The system fractionated most proteins according to molecular mass, although chemical reduction was required since certain proteins (e.g., bovine serum albumin) showed reduction-dependent shifts in mobility. Sodium dodecyl sulfate-PAGE of partially purified membrane proteins can be adversely affected by protein aggregation and proteolysis. PAU-PAGE, which solubilized aqueous insoluble proteins and rapidly inactivated proteases, was useful for assessing the polypeptide composition of plasma membrane preparations.

摘要

通过采用平板凝胶形式简化了酚 - 乙酸 - 脲聚丙烯酰胺凝胶电泳系统(PAU - PAGE),可同时比较多达12个样品。该系统根据分子量分离大多数蛋白质,不过由于某些蛋白质(如牛血清白蛋白)在迁移率上呈现出依赖还原的变化,所以需要进行化学还原。部分纯化的膜蛋白的十二烷基硫酸钠 - PAGE可能会受到蛋白质聚集和蛋白水解的不利影响。PAU - PAGE能够溶解水不溶性蛋白质并快速使蛋白酶失活,有助于评估质膜制剂的多肽组成。

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