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量化和站点本地化两个翻译后修改的同步亲和力丰富

Journal JoVE
Biologie

This content is Free Access.

Journal JoVE Biologie

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

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量化和站点本地化两个翻译后修改的同步亲和力丰富

DOI:

10.3791/60780-v

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12:11 min

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February 27, 2020

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Xueshu Xie, Samah Shah, Anja Holtz, Jacob Rose, Nathan Basisty, Birgit Schilling

¹Buck Institute for Research on Aging

Chapitres

00:04Introduction
01:19Desalting of Non-Enriched Proteolytic Peptides via Large-Scale Solid-Phase Extraction
02:46Simultaneous Enrichment of K-acetylated and K-succinylated Peptides with Immunoaffinity Beads
04:30Elution of the Peptides Bound to Antibody Beads
07:02Data Acquisition Using DDA (Data-Dependent Acquisition) and DIA (Data-Independent Acquisition)
10:02Results: One-pot Enrichment of PTMs
11:19Conclusion

Summary

Traduction automatique

此工作流程描述了用于定量全球蛋白质组分析的多个蛋白质转化后修饰（PTM）的及时且经济高效的扩充性能。该协议利用肽级PTM富集与多个结合抗体，然后数据独立的采集质谱分析，以获得对PTM串扰的生物见解。

Tags

Keywords: Simultaneous Affinity Enrichment Post-translational Modifications Quantification Site Localization Mass Spectrometry Acetylation Succinylation Protein Modifications PTMs Reproducibility C18 Resin Immunoaffinity Purification

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