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生物化学
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LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics
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生物化学
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JoVE 杂志 生物化学
LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics

LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics

DOI:
10.3791/55626-v

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08:01 min

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April 09, 2017

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, ,
1Division of Structural Biology and Biochemistry, School of Biological Sciences,Nanyang Technological University

Chapters

  • 00:05Title
  • 00:44Long-length Anion-exchange (LAX) Capillary Column Packing
  • 01:43Tissue Sample Preparation
  • 05:33One-dimension Long-length Electrostatic Repulsion-hydrophilic Interaction Chromatography-tandem Mass Spectrometry (LERLIC-MS/MS) Separation and Data Analysis
  • 06:48Results: Separation of Deamidation Isoforms of Glutamine and Asparagine by LERLIC-MS/MS
  • 07:34Conclusion

Summary

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Here we present a step-by-step protocol of the long-length electrostatic repulsion-hydrophilic interaction chromatography-tandem mass spectrometry (LERLIC-MS/MS) method. This is a novel methodology that enables for the first time quantification and characterization of the glutamine and asparagine deamidation isoforms by shotgun proteomics.

Tags

Keywords: LERLIC-MS/MSGlutamine DeamidationAsparagine DeamidationShotgun ProteomicsPeptide SeparationAnion Exchange ChromatographyProtein QuantificationDithiothreitolIodoacetamide

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