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Biochemistry

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JoVE Journal Biochemistry
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

Article DOI: 10.3791/63735-v • 09:16 min • May 4th, 2022
May 4th, 2022 •
1, 1, 1,2
1Department of Chemistry, University of Wisconsin-Madison, 2School of Pharmacy, University of Wisconsin-Madison

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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.

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