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.
Figure 1. Sandwich ELISA analysis of complex glycans
Materials and Reagents
Equipment
Procedure
Notes
Recipes
Acknowledgments
The research leading to this protocol has received funding from the European Union Seventh Framework Programme (FP7 2007-2013) under the WallTraC project (Grant Agreement n 263916). This article reflects the author’s views only. The European Community is not liable for any use that may be made of the information contained herein.
References
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