化学
组氨酸
重组DNA
咪唑
色谱法
洗脱
亲和层析
蛋白质纯化
串联亲和纯化
生物化学
组合化学
酶
基因
作者
Joanne Crowe,Brigitte Steude Masone,Joachim Ribbe
出处
期刊:Humana Press eBooks
[Humana Press]
日期:2003-11-14
卷期号:: 491-510
被引量:13
标识
DOI:10.1385/0-89603-402-x:491
摘要
The 6xHis/Ni-NTA system is a fast and versatile tool for the affinity purification of recombinant proteins and antigenic peptides. It is based on the high-affinity binding of six consecutive histidine residues (the 6xHis tag) to immobilized nickel ions, giving a highly selective interaction that allows purification of tagged proteins or protein complexes from <1% to >95% homogeneity in just one step (,). The tight association between the tag and the Ni-NTA resin allows contaminants to be easily washed away under stringent conditions, yet the bound proteins can be gently eluted by competition with imidazole, or a slight reduction in pH. Moreover, because the interaction is independent of the tertiary structure of the tag, 6xHis-tagged proteins can be purified even under the strongly denaturing conditions required to solubilize inclusion bodies.
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