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Mejora En-gel β-eliminación para ligada al O y exhaustiva sulfo-glycomics reductoras de Espectrometría de Masas
JoVE Journal
Chemistry
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JoVE Journal Chemistry
Improved In-gel Reductive β-Elimination for Comprehensive O-linked and Sulfo-glycomics by Mass Spectrometry
DOI:

13:06 min

November 20, 2014

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Chapters

  • 00:05Title
  • 01:16Gel Excision and Removal of Gel-derived Contaminants
  • 03:14In-gel O-glycan Release by Reductive β-Elimination and Desalting on Cation-exchange Chromatography
  • 06:02Borate Removal, C18 Clean-up and Base Preparation for Permethylation
  • 08:03Permethylation of Released Glycans C18 Clean-up and Mass Spectrometry Analysis
  • 10:33Results: Recovery and Mass Spectral Analysis of Permethylated O-linked Glycans
  • 12:30Conclusion

Summary

Automatic Translation

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.

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