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Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases
JoVE Journal
Biologia
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JoVE Journal Biologia
Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases

Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases

DOI:
10.3791/3749-v

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09:54 min

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December 26, 2011

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, ,
1New England Biolabs

Capitoli

  • 00:05Titolo
  • 01:46Enzymatic Deglycosylation
  • 03:32SDS-PAGE of Deglycosylated Samples
  • 04:55Pro-Q Emerald 300 for Detection of Glyclosylated Proteins in SDS-PAGE Gels
  • 06:49Analysis of hCGβ Enzymatic Deglycosylation
  • 09:02Conclusion

Summary

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Using specific glycosidases to remove sugars from glycoproteins followed by SDS-PAGE is a valuable method to detect glycan modifications on protein samples and is a good choice for initial glycobiology studies. Changes following deglycosylation can be detected as shifts in gel mobility or by staining with glycan sensitive reagents.

Tags

Protein GlycosylationSpecific EndoglycosidasesSpecific ExoglycosidasesPost-translational ModificationCell AdhesionMolecular TraffickingClearanceSignal TransductionN-linked GlycosylationO-linked GlycosylationHigh Mannose N-glycansComplex N-glycansO-glycansN-Acetylgalactosamine (GalNAc)GlycosidasesGlycosylation AnalysisRecombinant Human Chorionic Gonadotropin Beta (hCGβ)

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