1. A new monoclonal antibody against rhamnogalacturonan II and its application to immunocytochemical detection of rhamnogalacturonan II in Arabidopsis roots
- Author
-
Tatsuya Awano, Masaru Kobayashi, Keiji Takabe, Toru Matoh, and Ye Zhou
- Subjects
0106 biological sciences ,0301 basic medicine ,Rhamnogalacturonan-II ,food.ingredient ,Pectin ,medicine.drug_class ,Arabidopsis ,Enzyme-Linked Immunosorbent Assay ,Monoclonal antibody ,Plant Roots ,01 natural sciences ,Applied Microbiology and Biotechnology ,Biochemistry ,Analytical Chemistry ,Cell wall ,03 medical and health sciences ,chemistry.chemical_compound ,food ,Freezing ,Pressure ,medicine ,Arabidopsis thaliana ,Microscopy, Immunoelectron ,Molecular Biology ,biology ,Chemistry ,Organic Chemistry ,Antibodies, Monoclonal ,General Medicine ,Cell plate ,Chromatography, Ion Exchange ,biology.organism_classification ,Immunohistochemistry ,030104 developmental biology ,Solubilization ,Pectins ,010606 plant biology & botany ,Biotechnology - Abstract
Rhamnogalacturonan II (RG-II) is a region of pectin macromolecules that is present in plant primary cell walls. RG-II can be solubilized from cell walls as a borate-RG-II complex (B-RG-II), where two RG-II fragments are cross-linked via a borate diester linkage. Here, a rabbit monoclonal antibody against B-RG-II was prepared, which recognized both B-RG-II and RG-II monomers without borate ester-crosslinking. A pectic fragment with unknown structure was also recognized by the antibody, but neither homogalacturonan nor rhamnogalacturonan I was recognized. Immunoelectron microscopic analyses of Arabidopsis root tip cells were performed using this antibody. The signal was detected in developing cell plates and cell walls, which were denser in longitudinal walls than in transverse walls. These results coincide with our previous results obtained in suspension cultured tobacco cells, confirming that RG-II is present in cell plates at an early stage of their assembly. Abbreviations: B: boron; B-RG-II: borate-RG-II complex; ELISA: enzyme-linked immunosorbent assay; IgG: immunoglobulin G; mBSA: methylated bovine serum albumin; PGA: polygalacturonic acid; PLL: poly-l-lysine; RG-I: rhamnogalacturonan I; RG-II: rhamnogalacturonan II
- Published
- 2018