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Updated: Aug 17, 2025

Fluorescent Immunolocalization of Arabinogalactan Proteins and Pectins in the Cell Wall of Plant Tissues
Published on: February 27, 2021
Microscopy and Immunocytochemistry-Based Methods to Study Cell Wall Biosynthetic Enzymes in the Golgi
Sang-Jin Kim1,2,3, Federica Brandizzi4,5,6
1Department of Plant Biology, Michigan State University, East Lansing, MI, USA.
Abstract:
The Golgi apparatus has essential roles in all eukaryotic cells, and its importance in plants is further exemplified by a critical role in building a cellulosic cell wall. The Golgi apparatus houses numerous cell wall-synthesizing or cell wall-modifying enzymes to generate the complex cell wall structure. However, several putative cell wall biosynthetic candidates await characterization, which requires verification of the subcellular localization and enzymatic products. Here, we describe detailed methods to analyze the localization of proteins that are transiently produced in tobacco leaves or stably produced in transgenic plants, by confocal microscopy using fluorescent-tagged proteins along with known Golgi markers or the trafficking inhibitor brefeldin A. We also detail a procedure to analyze the enzymatic products through antibody-based immunoblotting after cell wall enrichment.
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