title: Improved sample preparation for CE-LIF analysis of plant N-glycans.
abstract: 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.
authors.affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
authors.forename: Bieke
authors.initials: B
authors.lastname: Nagels
authors.forename: Francis
authors.initials: F
authors.lastname: Santens
authors.forename: Koen
authors.initials: K
authors.lastname: Weterings
authors.forename: Els J M
authors.initials: EJ
authors.lastname: Van Damme
authors.forename: Nico
authors.initials: N
authors.lastname: Callewaert
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry*; D010944:Plants / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D014357:Trypsin / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D010446:Peptide Fragments; D010940:Plant Proteins; D011134:Polysaccharides; D014357:Trypsin