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用于蛋白质分析的十二烷基硫酸钠琼脂糖凝胶

SDS agarose gels for analysis of proteins.

作者信息

Wu M, Kusukawa N

机构信息

FMC BioProducts, Rockland, ME, USA.

出版信息

Biotechniques. 1998 Apr;24(4):676-8. doi: 10.2144/98244pf01.

Abstract

A new agarose-based protein electrophoresis gel system is described. The system consists of a highly resolving agarose, MetaPhor XR (FMC BioProducts, Rockland, ME, USA) dissolved in urea and TBE buffer and a stacking gel composed of a high gel-strength agarose, SeaKem Gold (FMC BioProducts). TBE containing sodium dodecyl sulfate (SDS) is used as electrophoresis buffer. The disadvantages of traditional agarose gels have been overcome, and several advantages over polyacrylamide gels have been demonstrated. The system is capable of high-resolution separation of small proteins and has a dynamic separation range equivalent to a 4%-20% gradient polyacrylamide gel. Furthermore, the staining of protein bands by Coomassie Brilliant Blue is very uniform in this gel, and depending on the protein, higher detection sensitivity can be obtained compared to SDS polyacrylamide gels. In Western blotting, proteins are more efficiently transferred to the membrane from the agarose gel than from polyacrylamide gels. Finally, the exceptional stability of agarose allows for gels to be precast and stored for a year.

摘要

本文描述了一种新型的基于琼脂糖的蛋白质电泳凝胶系统。该系统由溶解于尿素和TBE缓冲液中的高分辨率琼脂糖MetaPhor XR(美国缅因州罗克兰市FMC生物制品公司)以及由高凝胶强度琼脂糖SeaKem Gold(FMC生物制品公司)组成的堆积凝胶构成。含有十二烷基硫酸钠(SDS)的TBE用作电泳缓冲液。传统琼脂糖凝胶的缺点已被克服,并且已证明其相对于聚丙烯酰胺凝胶具有若干优点。该系统能够对小蛋白质进行高分辨率分离,其动态分离范围相当于4%-20%梯度聚丙烯酰胺凝胶。此外,考马斯亮蓝对该凝胶中蛋白质条带的染色非常均匀,并且根据蛋白质的不同,与SDS聚丙烯酰胺凝胶相比可获得更高的检测灵敏度。在蛋白质印迹法中,蛋白质从琼脂糖凝胶转移到膜上比从聚丙烯酰胺凝胶转移更有效。最后,琼脂糖的卓越稳定性使得凝胶可以预制并储存一年。

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