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Biochemistry
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用等压标记、广泛的液相色谱法、质谱法和软件辅助定量技术进行深层蛋白质组分析
JoVE Journal
Biochemistry
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JoVE Journal
Biochemistry
Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Please note that all translations are automatically generated.
Click here for the English version.
用等压标记、广泛的液相色谱法、质谱法和软件辅助定量技术进行深层蛋白质组分析
DOI:
10.3791/56474-v
•
10:37 min
•
November 15, 2017
•
Anthony A. High
,
Haiyan Tan
,
Vishwajeeth R. Pagala
,
Mingming Niu
3
,
Ji-Hoon Cho
,
Xusheng Wang
,
Bing Bai
,
Junmin Peng
2
1
St. Jude Proteomics Facility
,
St. Jude Children’s Research Hospital
,
2
Departments of Structural Biology and Developmental Neurobiology
,
St. Jude Children’s Research Hospital
,
3
Heilongjiang University of Chinese Medicine
Chapters
00:05
Title
01:00
Preparation of Cells/Tissues
01:45
Protein Extraction, Quality Control Western Blotting, In-solution Digestion, and Peptide Desalting
04:45
TMT Labeling of Peptides
05:50
Extensive High-resolution, Basic pH LC Prefactionation
06:43
LC-MS/MS Preparation and Parameters
07:41
Results: Deep Proteome Profiling Analysis
08:45
Conclusion
Summary
Automatic Translation
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Automatic Translation
我们提出了一个协议, 以准确定量蛋白质与等压标签, 广泛的分馏, 生物信息学工具, 和质量控制步骤, 结合液相色谱与高分辨率质谱仪接口。
Tags
Proteome Profiling
Isobaric Labeling
Liquid Chromatography
Mass Spectrometry
Quantification
Protein Identification
Protein Quantitation
Biomarker Discovery
Cell Lysis
Protein Digestion
Acetonitrile
Lys-C Protease
DTT
HEPES
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