Affinity Purification of Soluble and Membrane-Bound Protein Complexes by a FlpIn Strategy.

Autor:	Peschel R; Abteilung für Systembiochemie, Medizinische Fakultät der Ruhr-Universität Bochum, Bochum, Germany., Schmidt N; Abteilung für Systembiochemie, Medizinische Fakultät der Ruhr-Universität Bochum, Bochum, Germany., Schliebs W; Abteilung für Systembiochemie, Medizinische Fakultät der Ruhr-Universität Bochum, Bochum, Germany., Erdmann R; Abteilung für Systembiochemie, Medizinische Fakultät der Ruhr-Universität Bochum, Bochum, Germany.; Institut für Biochemie und Pathobiochemie, Ruhr-Universität Bochum, Bochum, Germany.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2643, pp. 373-382.
DOI:	10.1007/978-1-0716-3048-8_26
Abstrakt:	For a long time, the isolation of native protein complexes from human cells was accomplished by immunoprecipitation experiments. However, success depends on the quality of the antibodies and the method consumes valuable antibodies, which can hinder subsequent analysis of the isolated complexes. Here, we demonstrate an alternative approach based on affinity purification. It utilizes human Flp-In TM  cells, which genomically express a Protein A-tagged version of the human peroxisomal import receptor PEX5L. Native soluble and membrane-bound complexes containing PEX5L can thereby be isolated via a well-known affinity-based strategy. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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