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定量蛋白质组学使用还原二甲基化的稳定同位素标记

JoVE Journal
Química

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JoVE Journal Química

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

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定量蛋白质组学使用还原二甲基化的稳定同位素标记

DOI:

10.3791/51416-v

•

11:53 min

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July 01, 2014

•

Andrew C. Tolonen^2,3, Wilhelm Haas

¹CEA, DSV, IG,Genoscope, ²CNRS-UMR8030, Évry, France, ³Université d’Évry Val d’Essonne, ⁴Massachusetts General Hospital Cancer Center

Capítulos

00:05Título
01:19Protein Purification and Alkylation of Free Sulfhydryl Groups
02:59Protein Digestion, Reversed Phase Peptide Extraction, and Reductive Dimethylation Labeling
06:02Basic pH Reversed Phase Chromatography and STop and Go Extraction
08:17Microcapillary LC-MS/MS and Peptide Quantification
09:58Results: Accuracy, Precision, and Reproducibility of ReDi Labeling
11:24Conclusion

Summary

Tadução automática

肽的还原性二甲基化（REDI标签）稳定同位素标记是一种快速，廉价的战略精确质谱为基础的定量蛋白质组学。这里我们展示了一个可靠的方法用于制备使用可以应用于几乎任何类型的样品中的Redi途径的蛋白质混合物的分析。

Tags

Quantitative Proteomics Reductive Dimethylation Stable Isotope Labeling Mass Spectrometry Protein Quantification Peptide Labeling LC-MS/MS BPRP Chromatography StageTip Purification

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