Sandwich enzyme-linked immunosorbent assay (ELISA) analysis of plant cell wall glycan connections
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Date
2014-04-20
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2014 The Authors; exclusive licensee Bio-protocol LLC.
Abstract
Sandwich ELISA is a highly sensitive method that can be used to determine if two epitopes are part of the same macromolecule or supramolecular complex. In the case of plant cell wall glycans, it can reveal the existence of inter-polymers linkages, leading to better understanding of overall cell wall architectures. This development of a conventional sandwich ELISA protocol uses a carbohydrate-binding module (CBM), a small protein domain found in some carbohydrate catalysing or activating enzymes, and rat monoclonal antibodies (mAbs) which can be combined in the same ELISA plate without risk of cross reaction; the secondary anti-rat HRP antibody being only able to bind to the rat mAb and not the CBM. This protocol was developed and modified in the Prof. J. Paul Knox lab at the University of Leeds
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Keywords
ELISA, Polysaccharide, Carbohydrate, Plant biochemistry
Citation
2014