Biochemical and immunocytochemical characterization of monoclonal antibody TOP 35 raised against purified tonoplast from cress root

The vacuolar membrane, the tonoplast, is a proteinrich membrane hitherto only few proteins in it have been identified. As an approach for the identification of tonoplast proteins by monoclonal antibodies (MABs), purified tonoplast from cress roots (Lepidium sativum L.) were used for immunization and...

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Bibliographic Details
Published in:Journal of experimental botany 1997-06, Vol.48 (6), p.1215-1221
Main Authors: Liedtke, Christa, Polsakiewicz, Monika, Hartmann, Ingrid, Peters, Petra, Volkmann, Dieter, Scherer, G¨nther F.E.
Format: Article
Language:English
Subjects:
Antibodies
Antigens
Biochemistry
Biological and medical sciences
Cell and Molecular Biology and Biochemistry
Cell biochemistry
Cell membranes
Cell physiology
Electrophoresis
Fundamental and applied biological sciences. Psychology
Gels
Immnocytochemistry
Lepidium sativum
Monoclonal antibodies
monoclonal antibody
Plant antigens
Plant physiology and development
secretion
Tonoplast
vacuole
Vacuoles
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Summary:The vacuolar membrane, the tonoplast, is a proteinrich membrane hitherto only few proteins in it have been identified. As an approach for the identification of tonoplast proteins by monoclonal antibodies (MABs), purified tonoplast from cress roots (Lepidium sativum L.) were used for immunization and plasma membranes as a control membrane to test the absence of antigen. The MAB TOP 35 identified a glycoprotein of about 35 kDa in purified tonoplast of cress roots. Triton X-114 phase separation showed that it was a hydrophobic integral membrane protein. In immunocytochemistry the MAB TOP 35 strongly labelled the vacuolar membrane. The absence of cell wall or plasma membrane labelling by TOP 35 indicates a distinct biosynthetic pathway of this protein to the vacuolar membrane in plants.
ISSN:0022-0957
1460-2431
DOI:10.1093/jxb/48.6.1215