Immunofluorescent localization of MAPKs and colocalization with microtubules in Arabidopsis seedling whole-mount probes
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Arabidopsis cytology enzymology growth & development metabolism MeSH
- Fluorescent Antibody Technique methods MeSH
- Microscopy, Confocal MeSH
- Microtubules metabolism MeSH
- Mitogen-Activated Protein Kinases analysis metabolism MeSH
- Seedlings cytology enzymology growth & development MeSH
- Protein Transport MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Mitogen-Activated Protein Kinases MeSH
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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