Use of thiourea to increase the solubility of membrane proteins in two-dimensional electrophoresis
Use of thiourea to increase the solubility of membrane proteins in two-dimensional electrophoresis
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DOI：
10.1002/elps.1150190526
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发表时间：
1998-05-01
期刊：
影响因子：
2.9
通讯作者：
Rabilloud, T
中科院分区：
文献类型：
--
作者：
Rabilloud, T
采用高分辨率双向电泳法对膜蛋白进行了分离。在高负荷时，这些蛋白质容易沉淀，导致分辨率较低。这表明，硫脲的使用，先前描述的用于在固定的pH梯度中聚焦，可以扩展到传统的等电聚焦。由于硫脲对丙烯酰胺聚合有抑制作用，因此必须采用改进的光聚合体系。这些修饰提高了离子交换过程中蛋白质的溶解度，从而提高了二维凝胶的分辨率和容量。
The separation of membrane proteins by high-resolution two-dimensional electrophoresis was carried out. At high loads, these proteins are prone to precipitation, resulting in poor resolution. It is shown here that the use of thiourea, previously described for focusing in immobilized pH gradients, can be extended to conventional isoelectric focusing. As thiourea inhibits acrylamide polymerization, a modified photopolymerization system must be used. These modifications result in higher solubility of proteins during IEF, thereby increasing the resolution and capacity of the two-dimensional gels.