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N -连接糖链的糖蛋白在哺乳动物细胞中的脉冲大通分析
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JoVE Journal
Biology
Pulse-chase Analysis of N-linked Sugar Chains from Glycoproteins in Mammalian Cells
Please note that all translations are automatically generated.
Click here for the English version.
N -连接糖链的糖蛋白在哺乳动物细胞中的脉冲大通分析
DOI:
10.3791/1899-v
•
10:17 min
•
April 27, 2010
•
Edward Avezov
,
Efrat Ron
,
Yana Izenshtein
,
Yosef Adan
,
Gerardo Z. Lederkremer
1
Department of Cell Research and Immunology, George Wise Faculty of Life Sciences
,
Tel Aviv University
Chapters
00:00
Title
00:48
Introduction
01:28
Pulse-chase Labeling and Protein Extraction
04:34
Immunoprecipitation (IP)
05:38
Deglycosylation and Preparing the Samples for HPLCE Analysis
08:05
Analysis of N-glycans from NIH 3T3 Glycoproteins Representative Results
09:24
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
我们描述了一种通过改变早期的生活后,其在哺乳动物细胞中合成的糖蛋白的N -连接聚糖分析方法。这是通过分析代谢标记聚糖,从糖蛋白和高效液相色谱法检查酶的释放脉冲追逐。
Tags
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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