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Related Experiment Videos

Quantitative proteomics employing primary amine affinity tags.

Van M Hoang1, Thomas P Conrads, Timothy D Veenstra

  • 1Protein Chemistry Laboratory, National Cancer Institute at Frederick, SAIC-Frederick, Frederick, MD 21702-1201, USA.

Journal of Biomolecular Techniques : JBT
|September 19, 2003
PubMed
Summary
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This study introduces a novel proteomics method for quantifying protein levels using stable isotope labeling. The technique effectively measures relative protein abundance, offering a valuable tool for proteomic research.

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Quantitative proteomics is crucial for understanding biological processes.
  • Accurate methods for measuring relative protein abundance are needed.
  • Stable isotope labeling offers a powerful approach for quantitative proteomics.

Purpose of the Study:

  • To describe a new proteomics-based method for assessing relative peptide or protein abundance.
  • To demonstrate the efficacy of stable isotope labeling in quantitative proteomics.

Main Methods:

  • Proteins (bradykinin, carbonic anhydrase) were labeled using sulfosuccinimidyl-2-(biotinamido) ethyl-1,3-dithiopropionate, a primary amine-reactive reagent.
  • Tandem mass spectrometry confirmed the label's specificity to primary amines.

Related Experiment Videos

  • Secondary labeling with methyl iodide (natural and stable isotope-labeled) enabled relative quantitation.

    • Main Results:

    • The labeling reagent demonstrated specificity for primary amines.
    • The method allowed for the relative quantitation of labeled peptides/proteins.
    • Successful application of stable isotope labeling for quantitative analysis was achieved.

    Conclusions:

    • The described method is an effective stable isotope-labeling technique for quantitative proteomics.
    • This approach provides a reliable means to determine relative protein abundance.
    • The findings contribute to advancements in quantitative proteomic methodologies.