Niepmann Michael
Justus-Liebig-University Giessen, Institute of Biochemistry, Faculty of Medicine, Giessen, Germany.
Expert Rev Proteomics. 2007 Jun;4(3):355-61. doi: 10.1586/14789450.4.3.355.
A new native protein gel system was recently developed that enables the rapid and convenient analysis of virtually all soluble proteins, in particular including basic proteins, in their native oligomeric states. This gel system combines the addition of negative charges to the proteins by the dye SERVA Blue G with a Tris-histidine discontinuous buffer system and the use of polyacrylamide gradient gels. The use of histidine for sample focusing rather than glycine as a slow dipolar ion following from the cathode buffer serves to improve migration of basic proteins. In this review, the principle of function as well as the advantages and disadvantages of the new gel system are discussed in the context of other native protein gel systems and further methods for the analysis of the oligomeric state of a protein.
最近开发了一种新的天然蛋白质凝胶系统,该系统能够快速、方便地分析几乎所有处于天然寡聚状态的可溶性蛋白质,特别是碱性蛋白质。这种凝胶系统将染料SERVA Blue G给蛋白质添加负电荷与Tris-组氨酸不连续缓冲系统以及聚丙烯酰胺梯度凝胶的使用相结合。使用组氨酸而不是甘氨酸作为阴极缓冲液之后的慢偶极离子进行样品聚焦,有助于改善碱性蛋白质的迁移。在这篇综述中,结合其他天然蛋白质凝胶系统以及蛋白质寡聚状态分析的进一步方法,讨论了新凝胶系统的功能原理以及优缺点。