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二维电泳中蛋白质增溶的进展

Advances in protein solubilisation for two-dimensional electrophoresis.

作者信息

Herbert B

机构信息

Proteome Systems Ltd., North Ryde, Sydney, Australia.

出版信息

Electrophoresis. 1999 Apr-May;20(4-5):660-3. doi: 10.1002/(SICI)1522-2683(19990101)20:4/5<660::AID-ELPS660>3.0.CO;2-Q.

Abstract

Two-dimensional (2-D) electrophoresis remains the highest resolution technique for protein separation and is the method of choice when complex samples need to be arrayed for characterisation, as in proteomics. However, in current proteome projects the total number of proteins identified from 2-D gels is often only a small percentage of the predicted proteome. In addition, there is an almost complete lack of hydrophobic proteins on 2-D gels, especially those using immobilised pH gradients. Recently there have been a number of publications reporting reagents which improve protein solubilisation prior to isoelectric focusing. The improved solubilization possible with these reagents has increased the total number of proteins able to be visualised on 2-D gels and also allowed the separation of hydrophobic proteins, such as integral membrane proteins.

摘要

二维(2-D)电泳仍然是蛋白质分离分辨率最高的技术,并且是蛋白质组学中需要对复杂样品进行排列以进行表征时的首选方法。然而,在当前的蛋白质组计划中,从二维凝胶中鉴定出的蛋白质总数通常仅占预测蛋白质组的一小部分。此外,二维凝胶上几乎完全缺乏疏水蛋白,尤其是那些使用固定化pH梯度的凝胶。最近有许多出版物报道了在等电聚焦之前可改善蛋白质溶解的试剂。这些试剂可能实现的更好的溶解效果增加了能够在二维凝胶上可视化的蛋白质总数,并且还能够分离疏水蛋白,例如整合膜蛋白。

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