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Improved sample preparation for CE‐LIF analysis of plant N‐glycans
Author(s) -
Nagels Bieke,
Santens Francis,
Weterings Koen,
Van Damme Els J. M.,
Callewaert Nico
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100268
Subject(s) - glycomics , glycan , plant species , sample preparation , plant tissue , protocol (science) , chemistry , chromatography , computational biology , plant cell , biology , biochemistry , botany , medicine , glycoprotein , alternative medicine , pathology , gene
In view of glycomics studies in plants, it is important to have sensitive tools that allow one to analyze and characterize the N‐glycans present on plant proteins in different species. Earlier methods combined plant‐based sample preparations with CE‐LIF N‐glycan analysis but suffered from background contaminations, often resulting in non‐reproducible results. This publication describes a reproducible and sensitive protocol for the preparation and analysis of plant N‐glycans, based on a combination of the ‘in‐gel release method’ and N‐glycan analysis on a multicapillary DNA sequencer. Our protocol makes it possible to analyze plant N‐glycans starting from low amounts of plant material with highly reproducible results. The developed protocol was validated for different plant species and plant cells.