Abstract：
His-tagging is the most widespread and versatile strategy used to purify recombinant proteins for biochemical and structural studies. Recombinant DNA methods are first used to engineer the addition of a short tract of poly-histidine tag (His-tag) to the N-terminus or C-terminus of a target protein. The His-tag is then exploited to enable purification of the \"tagged\" protein by immobilized metal affinity chromatography (IMAC). In this chapter, we describe efficient procedures for the isolation of highly purified His-tagged target proteins from an Escherichia coli host using IMAC in a bind-wash-elute strategy that can be performed under both native and denaturing conditions.
摘要：
His标记是用于纯化重组蛋白以进行生物化学和结构研究的最广泛和通用的策略。重组DNA方法首先用于将短序列的多组氨酸标签(His标签)添加到靶蛋白的N-末端或C-末端。然后利用His标签，通过固定化金属亲和层析（IMAC）纯化“标记的”蛋白质。在这一章中,我们描述了在结合-洗涤-洗脱策略中使用IMAC从大肠杆菌宿主中分离高度纯化的His标记的靶蛋白的有效程序,该策略可以在天然和变性条件下进行.
