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Methods Enzymol. 2015;559:53-69. doi: 10.1016/bs.mie.2014.11.008. Epub 2015 May 16.

Strep-Tagged Protein Purification.

Author information

1
QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
2
QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany. Electronic address: frank.schaefer@qiagen.com.

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).

KEYWORDS:

Baculovirus-infected insect cells; E. coli lysates; Microscale purification; Strep-Tactin magnetic beads; Strep-tag system; Strep-tagged protein purification; Transfected mammalian cells

PMID:
26096503
DOI:
10.1016/bs.mie.2014.11.008
[Indexed for MEDLINE]

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