| Literature DB >> 26657801 |
Cebrail Karakus1, Merve Uslu2, Duygu Yazici2, Barik A Salih2.
Abstract
Immobilized metal affinity chromatography (IMAC) technique is used for fast and reliable purification of histidine(His)-tagged recombinant proteins. The technique provides purification under native and denaturing conditions. The aim of this study is to evaluate three commercially available IMAC kits (Thermo Scientific, GE Healthcare and Qiagen) for the purification of a 6xHis-tagged recombinant CagA (cytotoxin-associated gene A) protein from IPTG-induced Escherichia coli BL21(DE3) culture. The kits were tested according to the manufacturer instructions and the protein was purified with only GE Healthcare and Qiagen kits under denaturing conditions. 1% (w/v) SDS was used as denaturing agent in PBS instead of extraction reagent of Thermo Scientific kit to lyse bacterial cells from 100ml culture. The 6xHis-tagged recombinant protein was purified by the three kits equally.Entities:
Keywords: 6xHis-tag; Denaturing conditions; IMAC; Immobilized metal affinity chromatography; Protein purification; Recombinant protein; SDS
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Year: 2015 PMID： 26657801 DOI： 10.1016/j.jchromb.2015.11.045
Source DB: PubMed Journal: J Chromatogr B Analyt Technol Biomed Life Sci ISSN： 1570-0232 Impact factor: 3.205