Abstract
Tandem affinity purification (TAP) coupled to mass spectrometry has become a powerful approach to identify protein–protein interactions from different biological systems, including plants, in a proteome-wide manner. By using two sequential affinity purification steps, TAP allows for isolation of high-purity TAP-tagged proteins of interest and their associated proteins. Here we describe optimized procedures to use the GSRhino TAP technology for protein complex isolation from Arabidopsis cell suspension cultures.
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Acknowledgments
Research in our lab was supported by grants BIO2013-46539-R and BIO2016-80551-R funded by MINECO and AEI/FEDER/EU. M.G.-L. and E.I. were supported by FPI fellowships (Grant from “Severo Ochoa” Programme to M.G.-L.). We are very thankful to Geert de Jaeger, Jelle Van Leene, Geert Persiau and Dominique Eeckhout (VIB-Ghent, Belgium) for their help while setting up the TAP technique in our lab. We also acknowledge the CNB Proteomics facility for comments on the MS procedures.
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García-León, M., Iniesto, E., Rubio, V. (2018). Tandem Affinity Purification of Protein Complexes from Arabidopsis Cell Cultures. In: Oñate-Sánchez, L. (eds) Two-Hybrid Systems. Methods in Molecular Biology, vol 1794. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7871-7_21
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DOI: https://doi.org/10.1007/978-1-4939-7871-7_21
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