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Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
JoVE Journal
Biologia
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JoVE Journal Biologia
Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
DOI:

09:43 min

February 20, 2013

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Capitoli

  • 00:05Titolo
  • 01:43PALeO-TimeCourse: Protease-catalyzed Labeling of Proteolytic Cleavages
  • 02:43PALeO-TimeCourse: Postdigestion Labeling of Proteolytic Termini
  • 03:48ALeO-TimeCourse: Acid-catalyzed Labeling of Carboxyl Groups
  • 04:25MALDI-TOF/TOF MS/MS Data Acquisition and Analysis
  • 06:35Preparation of Spectral Time and 18O-incorporation Plots
  • 07:06Representative Results
  • 08:58Conclusion

Summary

Traduzione automatica

Stable isotope labeling workflows employing 18O-enriched water (LeO-workflows) are versatile tools for quantitative and qualitative proteomics studies. In protease-assisted (PALeO) workflows, 18O-atoms are introduced by proteolytic cleavage and carboxyl oxygen exchange reactions mediated by proteases. In the acid-catalyzed (ALeO) workflow, 18O-atoms are introduced by carboxyl oxygen exchange at low pH.

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