Summary
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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Acknowledgments
The authors thank Gabriele Dütsch, Elöd Körtvély, and Pia Jores for critical reading of the manuscript. This work was supported by the German Federal Ministry for Education and Research BMBF grant 031U108A/031U208A, BMBF grant 0316865A (QuantPro), and EU grants INTERACTION PROTEOME (LSHG-CT-2003-505520) and ProteomeBinders (FP6-026008). The authors C.J Gloeckner and K. Boldt contributed equally to this work.
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Gloeckner, C.J., Boldt, K., Schumacher, A., Ueffing, M. (2009). Tandem Affinity Purification of Protein Complexes from Mammalian Cells by the Strep/FLAG (SF)-TAP Tag. In: Reinders, J., Sickmann, A. (eds) Proteomics. Methods in Molecular Biology™, vol 564. Humana Press. https://doi.org/10.1007/978-1-60761-157-8_21
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DOI: https://doi.org/10.1007/978-1-60761-157-8_21
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