Targeted Purification of SnAvi-Tagged Proteins
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Tandem affinity purification (TAP) is a powerful technique to identify protein complex members. The modular composition of TAP-tags allows two sequential protein enrichment steps and thereby drastically reduces the amount of contaminants. Here, we describe the application of the SnAvi-tag—a TAP-tag useful in different expression systems. Due to its modular composition, this tag is multifunctional and facilitates among others the in vivo visualization of tagged proteins and their cell type specific activation.
Key wordsProtein complex purification Protein tag Tandem affinity purification (TAP) Tissue specificity Targeted in vivo biotinylation Tag activation In vivo visualization Mass spectrometry Protein interaction Protein modification
We would like to thank Angelika Schäfer, Antje Thien, Meta Rath, Caroline Scherzinger, Birte Manßhard, Caroline Keck, Gregor Bochenek, Erika v. Gromoff, and Birgit Holzwarth for experimental contributions to this project and Bettina Schulze for critical reading of the manuscript. Worm strains were obtained from the Caenorhabditis Genetics Center (CGC) at the University of Minnesota. The SB1 hybridoma cell line was developed by Michael Nonet and Gayla Hadwiger. We obtained it from the Developmental Studies Hybridoma Bank which was developed under the auspices of the NICHD and is maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242. This work was supported by Deutsche Forschungsgemeinschaft (CRC746).
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