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Chapters
- 00:00Title
- 00:48Introduction
- 01:28Pulse-chase Labeling and Protein Extraction
- 04:34Immunoprecipitation (IP)
- 05:38Deglycosylation and Preparing the Samples for HPLCE Analysis
- 08:05Analysis of N-glycans from NIH 3T3 Glycoproteins Representative Results
- 09:24Conclusion
Nós descrevemos um método para análise da alteração da N-glicanos ligados através do início da vida de glicoproteínas após a sua biossíntese em células de mamíferos. Isto é conseguido através de pulso-chase análise de glicanos metabolicamente marcados, a liberação enzimática das glicoproteínas e análise por HPLC.
Tags
Pulse-chase AnalysisN-linked Sugar ChainsGlycoproteinsMammalian CellsGlc3Man9GlcNAc2 Precursor OligosaccharideER ModificationSecretory ProteinsGlycoprotein Quality ControlTrafficking SignalsProteomic AnalysisDynamics Of Sugar Chain ProcessingLabeling And Pulse-chase Analysis ProtocolsMetabolically Labeled N-linked Oligosaccharides[2-3H] Mannose LabelingDenaturing LysisEnzymatic ReleaseImmunoprecipitated ProteinGeneral Glycoprotein PoolEndo H TreatmentN-glycosidase F TreatmentMolecular FiltrationHPLC Analysis
