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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 신문
생화학

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JoVE 신문 생화학

TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis

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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

DOI:

10.3791/60970-v

•

07:44 min

•

June 08, 2020

•

Silveli Suzuki-Hatano, Ang-Chen Tsai, Audrey Daugherty, Christina A. Pacak

¹Department of Pediatrics,University of Florida

Chapters

00:04Introduction
00:31Protein Extraction
01:40Reducing/Alkylating Reagent Treatment
02:39Methanol/Chloroform Precipitation
04:20Protein Digestion
04:56Peptide Labeling
05:57Results: Representative Protein Quantification and Volcano Plot and Protein Analysis through Evolutionary Relationships (PANTHER) Analyses
07:04Conclusion

Summary

자동 번역

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

Keywords: 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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