title: Immunofluorescent localization of MAPKs and colocalization with microtubules in Arabidopsis seedling whole-mount probes.
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.
authors.affiliation: Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University Olomouc, Šlechtitelů 11, 783 71, Olomouc, Czech Republic, olga.samajova@upol.cz.
authors.forename: Olga
authors.initials: O
authors.lastname: Samajová
authors.forename: George
authors.initials: G
authors.lastname: Komis
authors.forename: Jozef
authors.initials: J
authors.lastname: Samaj
mesh_terms: D017360:Arabidopsis / Q000166:cytology* / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods*; D018613:Microscopy, Confocal; D008870:Microtubules / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000032:analysis / Q000378:metabolism*; D021381:Protein Transport; D036226:Seedlings / Q000166:cytology* / Q000201:enzymology* / Q000254:growth & development
chemical_list: D020928:Mitogen-Activated Protein Kinases