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Pulse-chase Analysis of N-linked Sugar Chains from Glycoproteins in Mammalian Cells
 

Pulse-chase Analysis of N-linked Sugar Chains from Glycoproteins in Mammalian Cells

Article DOI: 10.3791/1899-v • 10:18 min • April 27th, 2010
April 27th, 2010 •
1, 1, 1, 1, 1
1Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University

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We describe a method for analysis of the alteration of N-linked glycans through the early life of glycoproteins after their biosynthesis in mammalian cells. This is achieved by pulse-chase analysis of metabolically labeled glycans, enzymatic release from glycoproteins and examination by HPLC.

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Pulse-chase Analysis N-linked Sugar Chains Glycoproteins Mammalian Cells Glc3Man9GlcNAc2 Precursor Oligosaccharide ER Modification Secretory Proteins Glycoprotein Quality Control Trafficking Signals Proteomic Analysis Dynamics Of Sugar Chain Processing Labeling And Pulse-chase Analysis Protocols Metabolically Labeled N-linked Oligosaccharides [2-3H] Mannose Labeling Denaturing Lysis Enzymatic Release Immunoprecipitated Protein General Glycoprotein Pool Endo H Treatment N-glycosidase F Treatment Molecular Filtration HPLC Analysis
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