Abstract
Knowledge about the identity of the interacting partners is important for the understanding of the function and the cellular activity of a given protein. Here we describe co-immunoprecipitation and pull-down as methods that are widely used for the identification and characterization of protein–protein interactions. These methods are well suited to find or confirm the interaction among multiple proteins, given the availability of a specific antibody for or a tagged version of the protein of interest.
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References
Fields, S. and Song, O.-K. (1989) A novel genetic system to detect protein–protein interactions. Nature 340, 245–246.
Hu, C.-D., Chinenov, Y., and Kerppola, T. K. (2002) Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Mol Cell 9, 789–798.
Walter, M., Chaban, C., Schütze, K., Batistic, O., Weckermann, K., Näke C., Blazevic, D., Grefen, C., Schumacher, K., Oecking, C., Harter, K., and Kudla, J. (2004) Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation. Plant J 40, 428–438.
Kessler, S. W. (1975) Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunology 115, 1617–1624.
Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76, 4350–4354.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Willige, B. C., et al. (2007) The DELLA domain of GA INSENSITIVE mediates the interaction with the GA INSENSITIVE DWARF1A gibberellin receptor of Arabidopsis. Plant Cell 19, 1209–1220.
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Isono, E., Schwechheimer, C. (2010). Co-immunoprecipitation and Protein Blots. In: Hennig, L., Köhler, C. (eds) Plant Developmental Biology. Methods in Molecular Biology, vol 655. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-765-5_25
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DOI: https://doi.org/10.1007/978-1-60761-765-5_25
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-764-8
Online ISBN: 978-1-60761-765-5
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