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Immunofluorescent Localization of MAPKs in Steedman’s Wax Sections

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Plant MAP Kinases

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

Abstract

Signals of different nature are transduced in cells through signal transduction pathways, where mitogen-activated protein kinases (MAPKs) play an important role as signaling molecules. Views into intracellular localization of MAPKs are critical for the understanding of their spatial and temporal functions, like activation-based relocation, compartmentation, or interactions with local substrates. Localization of MAPKs in cells is thus very useful cell biological approach, extending complex mode of cell signaling characterization in plants. Here, we present a method for subcellular immunofluorescence localization of MAPKs using protein- or phospho-specific antibodies, performed on sectioned fixed plant samples. It is based on embedding of samples in the Steedman’s wax, a low-melting point polyester wax embedding medium, which maintains high antigenicity of studied proteins. In addition, exposure of dewaxed sections to antibodies allows for their efficient penetration. Altogether, it makes this simple method a good tool in the efficient subcellular localization of diverse proteins, including plant MAPKs.

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Acknowledgements

This work was supported by grant P501/11/1764 from the Czech Science Foundation (GAČR).

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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

    Miroslav Ovečka, Olga Šamajová & Jozef Šamaj

  2. Department of Plant Cell Biology, IZMB, University of Bonn, Kirschallee 1, 53115, Bonn, Germany

    František Baluška

Authors
  1. Miroslav Ovečka
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  2. Olga Šamajová
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  3. František Baluška
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  4. Jozef Šamaj
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Corresponding author

Correspondence to Miroslav Ovečka .

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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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Ovečka, M., Šamajová, O., Baluška, F., Šamaj, J. (2014). Immunofluorescent Localization of MAPKs in Steedman’s Wax Sections. 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_10

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  • DOI: https://doi.org/10.1007/978-1-4939-0922-3_10

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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

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