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一种同时分析人胰腺组织中N-糖肽和磷酸肽的自旋尖端富集策略

JoVE Journal
Biochimica

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JoVE Journal Biochimica

A Spin-Tip Enrichment Strategy for Simultaneous Analysis of N-Glycopeptides and Phosphopeptides from Human Pancreatic Tissues

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一种同时分析人胰腺组织中N-糖肽和磷酸肽的自旋尖端富集策略

DOI:

10.3791/63735-v

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09:16 min

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May 04, 2022

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Dylan Nicholas Tabang, Danqing Wang, Lingjun Li²

¹Department of Chemistry,University of Wisconsin-Madison, ²School of Pharmacy,University of Wisconsin-Madison

Capitoli

00:05Introduction
01:12Tissue Cryo-Pulverization, Lysis, Protein Extraction
03:35Protein Digestion and Desalting
05:36Dual-Functional Ti(IV) Simultaneous Enrichment
07:19Results: Dual-Functional Ti(IV)-IMAC Enrichment and Mass Spectrometry Analysis of Protein N-Glycosylation and Phosphorylation in Pancreatic Tissues
08:37Conclusion

Summary

Traduzione automatica

翻译后修饰（PTM）改变蛋白质结构和功能。同时富集多个PTM类型的方法可以最大限度地提高分析的覆盖率。我们提出了一个使用双功能Ti（IV）固定化金属亲和色谱法，然后进行质谱分析的方案，用于同时富集和分析胰腺组织中的蛋白质N-糖基化和磷酸化。

Tags

Keywords: N-glycopeptides Phosphopeptides Titanium Immobilized Metal Affinity Chromatography Dual-functional Enrichment Simultaneous Analysis Disease Biomarker Discovery Diabetes Cancer Spin Tips Centrifugation Lysis Buffer Protease Inhibitor Phosphatase Inhibitor Sonication Mass Spectrometry Analysis

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