Plant MAP Kinases pp 117-130

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

| Cite as

Immunofluorescent Localization of MAPKs in Steedman’s Wax Sections

  • Miroslav Ovečka
  • Olga Šamajová
  • František Baluška
  • Jozef Šamaj
Protocol

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.

Key words

Arabidopsis thalianaImmunofluorescence microscopy MAPK-specific antibody Medicago sativaOxidative stress Salt stress Sections Steedman’s wax 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Miroslav Ovečka
    • 1
  • Olga Šamajová
    • 1
  • František Baluška
    • 2
  • Jozef Šamaj
    • 1
  1. 1.Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of SciencePalacký University OlomoucOlomoucCzech Republic
  2. 2.Department of Plant Cell Biology, IZMBUniversity of BonnBonnGermany

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