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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
Tandem affinity purification of protein complexes from mammalian cells by the Strep/FLAG (SF)-TAP tag.
Methods Mol. Biol. 564, 359-372 (2009)
Isolation and dissection of native multiprotein complexes is a central theme in functional genomics. The development of the tandem affinity purification (TAP) tag has enabled efficient and large-scale purification of native protein complexes. The SF-TAP tag, a modified version of the TAP tag, allows a fast and straightforward purification of protein complexes from mammalian cells. It consists of a tandem Strep-tag II and a FLAG epitope (SF-TAP). The SF-TAP tag allows a native elution of protein complexes without proteolytic cleavage needed in the original TAP procedure. Besides the SF-TAP protocol, the principal idea of a pathway mapping by subsequent tagging of copurified proteins is demonstrated for the interactome of the MAPKKK Raf.
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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Herausgeber
Reinders, R.* ; Sickmann, A.*
Schlagwörter
Protein complexes; Tandem affinity tag; TAP; SF-TAP
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Konferenztitel
Proteomics : Methods and Protocols
Zeitschrift
Methods in Molecular Biology
Quellenangaben
Band: 564,
Seiten: 359-372
Reihe
Methods in Molecular Biology
Verlag
Springer
Verlagsort
Berlin [u.a.]
Begutachtungsstatus
Peer reviewed
Institut(e)
CF Metabolomics & Proteomics (CF-MPC)