title: Strep-Tagged Protein Purification.
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).
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Maertens
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Spriestersbach
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Kubicek
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany. Electronic address: frank.schaefer@qiagen.com.
authors.forename: Frank
authors.initials: F
authors.lastname: Schäfer
mesh_terms: D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D007313:Insecta; D008024:Ligands; D008280:Magnetics; D009842:Oligopeptides / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry
chemical_list: C099517:Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly; D008024:Ligands; D009842:Oligopeptides; D011506:Proteins; D011994:Recombinant Proteins; D012685:Sepharose; D019809:Streptavidin
keywords: Baculovirus-infected insect cells; E. coli lysates; Microscale purification; Strep-Tactin magnetic beads; Strep-tag system; Strep-tagged protein purification; Transfected mammalian cells