| Literature DB >> 26096503 |
Barbara Maertens1, Anne Spriestersbach1, Jan Kubicek1, Frank Schäfer2.
Abstract
The Strep-tag system can be used to purify recombinant proteins from any expression system. Here, protocols for lysis and affinity purification of Strep-tagged proteins from E. coli, baculovirus-infected insect cells, and transfected mammalian cells are given. Depending on the amount of Strep-tagged protein in the lysate, a protocol for batch binding and subsequent washing and eluting by gravity flow can be used. Agarose-based matrices with the coupled Strep-Tactin ligand are the resins of choice, with a binding capacity of up to 9 mg ml(-1). For purification of lower amounts of Strep-tagged proteins, the use of Strep-Tactin magnetic beads is suitable. In addition, Strep-tagged protein purification can also be automated using prepacked columns for FPLC or other liquid-handling chromatography instrumentation, but automated purification is not discussed in this protocol. The protocols described here can be regarded as an update of the Strep-Tag Protein Handbook (Qiagen, 2009).Entities:
Keywords: Baculovirus-infected insect cells; E. coli lysates; Microscale purification; Strep-Tactin magnetic beads; Strep-tag system; Strep-tagged protein purification; Transfected mammalian cells
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Year: 2015 PMID： 26096503 DOI： 10.1016/bs.mie.2014.11.008
Source DB: PubMed Journal: Methods Enzymol ISSN： 0076-6879 Impact factor: 1.600