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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
Bioquímica

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JoVE Journal Bioquímica

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

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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

DOI:

10.3791/56474-v

•

10:37 min

•

November 15, 2017

•

Anthony A. High, Haiyan Tan, Vishwajeeth R. Pagala, Mingming Niu³, Ji-Hoon Cho, Xusheng Wang, Bing Bai, Junmin Peng²

¹St. Jude Proteomics Facility,St. Jude Children’s Research Hospital, ²Departments of Structural Biology and Developmental Neurobiology,St. Jude Children’s Research Hospital, ³Heilongjiang University of Chinese Medicine

Capítulos

00:05Título
01:00Preparation of Cells/Tissues
01:45Protein Extraction, Quality Control Western Blotting, In-solution Digestion, and Peptide Desalting
04:45TMT Labeling of Peptides
05:50Extensive High-resolution, Basic pH LC Prefactionation
06:43LC-MS/MS Preparation and Parameters
07:41Results: Deep Proteome Profiling Analysis
08:45Conclusion

Summary

Tadução automática

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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