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Plant MAP Kinases pp 107–115Cite as

Immunofluorescent Localization of MAPKs and Colocalization with Microtubules in Arabidopsis Seedling Whole-Mount Probes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1171))

Abstract

In all eukaryotes, signaling by mitogen-activated protein kinase (MAPK) pathways plays a crucial role in signal transduction during regulation of cell growth, differentiation, proliferation as well as death and stress responses. In this chapter we describe a reliable method to immunolocalize MAPKs in roots of Arabidopsis thaliana by using whole-mount seedling probes. This method relies on quick and efficient chemical fixation, partial cell wall digestion, plasma membrane permeabilization, subsequent antibody incubation, and visualization by high-end confocal laser scanning microscopy (CLSM) performed on whole Arabidopsis seedlings. Protocols are provided for immunofluorescent localization of MPK3, MPK4, and MPK6, representing three major developmentally and stress-regulated MAPKs of Arabidopsis. In addition, protocols for colocalization of these MAPKs with microtubules are also provided.

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Acknowledgements

This research was supported by National Program for Sustainability I (grant no. LO1204) provided by the Czech Ministry of Education.

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Authors and Affiliations

  1. Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University Olomouc, Šlechtitelů 11, 783 71, Olomouc, Czech Republic

    Olga Šamajová, George Komis & Jozef Šamaj

Authors
  1. Olga Šamajová
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  2. George Komis
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  3. Jozef Šamaj
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Corresponding author

Correspondence to Olga Šamajová .

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Editors and Affiliations

  1. Palacky University in Olomouc, Olomouc, Czech Republic

    George Komis

  2. Faculty of Science, Palacky University in Olomouc, Olomouc, Czech Republic

    Jozef Šamaj

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Šamajová, O., Komis, G., Šamaj, J. (2014). Immunofluorescent Localization of MAPKs and Colocalization with Microtubules in Arabidopsis Seedling Whole-Mount Probes. In: Komis, G., Šamaj, J. (eds) Plant MAP Kinases. Methods in Molecular Biology, vol 1171. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0922-3_9

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  • DOI: https://doi.org/10.1007/978-1-4939-0922-3_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0921-6

  • Online ISBN: 978-1-4939-0922-3

  • eBook Packages: Springer Protocols

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