Abstract
There is a growing demand for methods that allow rapid and reliable in situ localization of proteins in plant cells. The immunocytochemistry protocol presented here can be used routinely to observe protein localization patterns in tissue sections of various plant species. This protocol is especially suitable for plant species with more-complex tissue architecture (such as maize, Zea mays), which makes it difficult to use an easier whole-mount procedure for protein localization. To facilitate the antibody-antigen reaction, it is necessary to include a wax-embedding and tissue-sectioning step. The protocol consists of the following procedures: chemical fixation of tissue, dehydration, wax embedding, sectioning, dewaxing, rehydration, blocking and antibody incubation. The detailed protocol, recommended controls and troubleshooting are presented here, along with examples of applications.
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Acknowledgements
We are grateful to F. Baluska, P. Knox, W. Michalke and K. Roberts for sharing material; we thank P. Brewer for critical reading of the manuscript and discussions. M.S., J.W. and J.F were supported by VolkswagenStiftung and the European Molecular Biology Organization Young Investigator Program; T.P. was supported by the Deutsche Forschungsgemeinschaft (SFB 446); and J.B. was supported by Grant Agency of Czech Republic (522/02/D137) and Ministry of Education of Czech Republic (MSM432100001).
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Paciorek, T., Sauer, M., Balla, J. et al. Immunocytochemical technique for protein localization in sections of plant tissues. Nat Protoc 1, 104–107 (2006). https://doi.org/10.1038/nprot.2006.16
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DOI: https://doi.org/10.1038/nprot.2006.16
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