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

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

DOI:
10.3791/63735-v

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09:16 min

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May 04, 2022

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

Capítulos

  • 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

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Tadução automática

Post-translational modifications (PTMs) change protein structures and functions. Methods for the simultaneous enrichment of multiple PTM types can maximize coverage in analyses. We present a protocol using dual-functional Ti(IV)-immobilized metal affinity chromatography followed by mass spectrometry for the simultaneous enrichment and analysis of protein N-glycosylation and phosphorylation in pancreatic tissues.

Tags

Keywords: N-glycopeptidesPhosphopeptidesTitanium Immobilized Metal Affinity ChromatographyDual-functional EnrichmentSimultaneous AnalysisDisease Biomarker DiscoveryDiabetesCancerSpin TipsCentrifugationLysis BufferProtease InhibitorPhosphatase InhibitorSonicationMass Spectrometry Analysis

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