Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Absolute quantification strategies in proteomics based on mass spectrometry.

Mark Brönstrup1

  • 1Sanofi-Aventis, Building G878, D-65926 Frankfurt, Germany. Mark.Broenstrup@aventis.com

Expert Review of Proteomics
|June 22, 2005
PubMed
Summary

Absolute quantification using mass spectrometry provides precise protein abundance measurements. This method, utilizing isotope-dilution and comparing reference peptides to endogenous ones, is ideal for biomarker studies and post-translational modification analysis.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Cystobactamid off-target profiling reveals favorable safety, superoxide reduction, and SCARB1 inhibition in eukaryotes.

npj drug discovery·2026
Same author

MexXY-OprM efflux pump mediates resistance to odilorhabdins in <i>Pseudomonas aeruginosa</i>.

Microbiology spectrum·2026
Same author

Bioresponsive pseudoGlucosinolates (psGSLs) Release Isothiocyanates (ITCs) in the Presence of Nitroreductases.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
Same author

An Imidazo[2,1-b][1,3,4]thiadiazole Derivative Inhibits the Virulence Factor α-Hemolysin by Blocking the Pullout of Its Stem Domain.

ChemMedChem·2026
Same author

Call for Papers: Artificial Intelligence for Next-generation Anti-infective Discovery.

ACS infectious diseases·2026
Same author

A novel class of oligoarylamide antibiotics defined by albicidins and cystobactamids.

Natural product reports·2025

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Quantitative proteomics requires accurate protein abundance determination.
  • Mass spectrometry (MS) based methods are crucial for protein quantification.
  • Absolute quantification offers precise measurement of protein levels.

Purpose of the Study:

  • To review MS experiments for absolute protein quantification.
  • To highlight the isotope-dilution strategy for accurate measurements.
  • To discuss the application and limitations of absolute quantification.

Main Methods:

  • Utilizes the isotope-dilution concept with synthetic, isotope-labeled reference peptides.
  • Quantification relies on comparing MS signal intensities of reference peptides to endogenous peptides.

Related Experiment Videos

  • Applicable to determining post-translational modification levels at specific residues.

    • Main Results:

    • Achieves low detection limits (subfemtomole levels) and a high dynamic range (approx. five orders of magnitude).
    • Demonstrates compatibility with various MS techniques and sample purification methods.
    • Reports quantification errors and coefficients of variation below 5% under ideal conditions.

    Conclusions:

    • Absolute quantification is suitable for studies requiring precise protein numbers, repetitive experiments, or detailed post-translational modification analysis.
    • Potential errors can arise from reference peptide selection and proteolysis efficiency.
    • Key applications include biomarker quantification, studying phosphorylation/ubiquitination, and analyzing protein interactions.