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Related Experiment Video

Updated: Jul 4, 2026

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
09:43

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water

Published on: February 20, 2013

18O labeling over a coffee break: a rapid strategy for quantitative proteomics.

Shama P Mirza1, Andrew S Greene, Michael Olivier

  • 1National Center for Proteomics Research, Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA. smirza@mcw.edu

Journal of Proteome Research
|May 31, 2008
PubMed
Summary

This study introduces a rapid 15-minute (18)O labeling method for proteomic quantification. This technique enhances efficiency for differential protein expression analysis using mass spectrometry.

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Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Biochemistry

Background:

  • Accurate differential protein expression measurement is crucial in proteomics.
  • Isotopic labeling, specifically the (18)O method, offers a simple, cost-effective approach for relative quantification.
  • Traditional (18)O labeling requires lengthy incubation periods, limiting its widespread adoption.

Purpose of the Study:

  • To develop a significantly faster (18)O labeling protocol for proteomic quantification.
  • To improve the efficiency and reduce the time required for enzyme-mediated oxygen substitution.
  • To demonstrate the applicability of the accelerated labeling method for accurate protein expression analysis.

Main Methods:

  • Utilized immobilized trypsin on micro-spin columns for sample digestion and (18)O labeling.

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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Related Experiment Videos

Last Updated: Jul 4, 2026

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water
09:43

Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water

Published on: February 20, 2013

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

  • Optimized the enzyme-mediated oxygen substitution process to achieve rapid labeling.
  • Validated the method using a mixture of four proteins and whole cell lysates from rat vascular endothelial cells.

    • Main Results:

    • Successfully reduced the (18)O labeling procedure to 15 minutes.
    • Achieved high labeling efficiency within the accelerated timeframe.
    • Demonstrated the accuracy and efficiency of the rapid labeling method in complex biological samples.

    Conclusions:

    • The 15-minute (18)O labeling protocol significantly enhances the speed of proteomic quantification.
    • This accelerated method overcomes the time limitations of traditional (18)O labeling.
    • The developed technique provides a simple, efficient, and accurate tool for differential protein expression studies.