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

  • Olga ŠamajováEmail author
  • George Komis
  • Jozef Šamaj
Part of the Methods in Molecular Biology book series (MIMB, volume 1171)


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.

Key words

Arabidopsis thaliana Mitogen activated protein kinase Root Whole mount Immunolocalization 



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


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of SciencePalacký University OlomoucOlomoucCzech Republic

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