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Biochemistry
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蛋白质组学设施提交和后续数据分析的TMT样品制备
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal
Biochemistry
TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis
Please note that all translations are automatically generated.
Click here for the English version.
蛋白质组学设施提交和后续数据分析的TMT样品制备
DOI:
10.3791/60970-v
•
07:44 min
•
June 08, 2020
•
Silveli Suzuki-Hatano
,
Ang-Chen Tsai
,
Audrey Daugherty
,
Christina A. Pacak
1
Department of Pediatrics
,
University of Florida
Chapters
00:04
Introduction
00:31
Protein Extraction
01:40
Reducing/Alkylating Reagent Treatment
02:39
Methanol/Chloroform Precipitation
04:20
Protein Digestion
04:56
Peptide Labeling
05:57
Results: Representative Protein Quantification and Volcano Plot and Protein Analysis through Evolutionary Relationships (PANTHER) Analyses
07:04
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
我们提出了优化的串联质量标签 (TMT) 标签协议,其中包括以下每个步骤的详细信息:蛋白质提取、定量、沉淀、消化、标记、提交到蛋白质组学设施以及数据分析。
Tags
TMT Sample Preparation
Proteomics
Protein Extraction
Labeling Efficiency
Reproducibility
Data Acquisition
CHAPS Lysis Buffer
Bead Disruptor
Sonication
Reducing/alkylating Reagent Treatment
TCEP
Iodoacetamide
Methanol/chloroform Precipitation
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