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Ishikawa Prefectural University

1 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Improved In-gel Reductive β-Elimination for Comprehensive O-linked and Sulfo-glycomics by Mass Spectrometry

David B. Nix^1,2, Tadahiro Kumagai¹, Toshihiko Katoh^1,3, Michael Tiemeyer^1,2, Kazuhiro Aoki¹

¹Complex Carbohydrate Research Center, University of Georgia, ²Department of Biochemistry and Molecular Biology, University of Georgia, ³Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University

In order to comprehensively explore the diversity of O-linked glycans, a new procedure for in-gel reductive β-elimination, combined with permethylation and a rapid phase-partition method, is applied to the analysis of O-linked glycans directly released from glycoproteins resolved by SDS-PAGE and amenable to subsequent glycomic analysis by mass spectrometry.