Immunocytochemical fluorescent in situ visualization of proteins in Arabidopsis.
The understanding of cellular and subcellular functions often relies on the ability to visualize proteins as close as possible to their endogenous locations. A number of immunocytochemical techniques have been developed to detect proteins in situ using specific antibodies raised against proteins of interest. Here, we describe in detail two protocols commonly, successfully employed in Arabidopsis research. The first allows for immunolocalization of proteins in whole-mount Arabidopsis roots without the need for physical sectioning. The second allows for immunolocalization of proteins on semi-thin microtome sections of wax-embedded swamples. This approach is particularly useful when sectioning of Arabidopsis roots or other thicker plant organs is required for immunolocalization. We provide step-by-step protocols with extensive troubleshooting for both the whole-mount and sectioning protocols. Furthermore, critical steps, advantages, and limitations of the two protocols described here are discussed.
Department of Forest Genetics and Plant Physiology, UPSC, Swedish University of Agricultural Sciences, Umeå, Sweden.
Yohann
Y
Boutté
Markus
M
Grebe
D017360:Arabidopsis / Q000166:cytology / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D008856:Microscopy, Fluorescence; D018517:Plant Roots / Q000166:cytology / Q000378:metabolism; D021381:Protein Transport; D014617:Vacuoles / Q000378:metabolism
D029681:Arabidopsis Proteins