琼脂糖
电泳
考马斯亮蓝
聚丙烯酰胺凝胶电泳
色谱法
蛋白质凝胶电泳
凝胶电泳
化学
琼脂糖凝胶电泳
二维凝胶电泳
分子量大小标记
核酸凝胶电泳
高分子
颜色标记
自由流电泳
染色
生物化学
生物
蛋白质组学
DNA
酶
基因
遗传学
作者
Tsutomu Arakawa,Masahiko Nakagawa,Yui Tomioka,Chiaki Sakuma,Cynthia Li,Tomomi Satō,Ryo Sato,Takahiro Shibata,Yasunori Kurosawa,Teruo Akuta
标识
DOI:10.1016/bs.mcb.2021.12.030
摘要
Electrophoresis is one of the most important analytical technologies for characterization of macromolecules and their interactions. Among them, native gel electrophoresis is used to analyze the macromolecules in the native structure. It differs in principle and information from those obtained by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) or blue native polyacrylamide gel electrophoresis (BN-PAGE). SDS-PAGE is carried out in the presence of strong denaturant, SDS, while BN-PAGE is done in the presence of negatively charged dye, e.g., Coomassie brilliant blue, G-250. Here, we describe native gel electrophoresis using agarose gel and a buffer at pH 6.1 composed of histidine and 2-(N-morpholino) ethanesulfonic acid. First, a protocol for vertical and horizontal formats of agarose native gel electrophoresis is described followed by different staining procedures. Then, various examples obtained using the developed procedure will be shown to demonstrate how the technology can be applied to specific cases and the advantages or caveats of the present technology.
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