Abstract
Protein complex purification represents a powerful approach to identify novel players in plant innate immunity. However, the identification of interacting protein partners within a natural context has been a challenge for researchers. In this chapter, we describe a method of immunoaffinity chromatography using purified, antibodies to isolate native protein complexes from wild-type tissue. We detail the antibody purification and immobilization steps in addition to the co-immunoprecipitation protocol. In addition, a method to prepare protein samples for mass spectroscopy analysis is described. This straightforward protocol has been used to isolate and identify novel components of Arabidopsis immunity-associated protein complexes.
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Acknowledgments
We thank Brett Phinney at UC Davis Genome Center Proteomics Core Facility for providing the in-gel trypsin digestion protocol.
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Elmore, J.M., Coaker, G. (2011). Biochemical Purification of Native Immune Protein Complexes. In: McDowell, J. (eds) Plant Immunity. Methods in Molecular Biology, vol 712. Humana Press. https://doi.org/10.1007/978-1-61737-998-7_4
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DOI: https://doi.org/10.1007/978-1-61737-998-7_4
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