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 Video

Updated: May 19, 2026

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
16:10

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

High-resolution enabled TMT 8-plexing.

Thilo Werner1, Isabelle Becher, Gavain Sweetman

  • 1Cellzome AG, Meyerhofstrasse 1, 69117 Heidelberg, Germany.

Analytical Chemistry
|August 14, 2012
PubMed
Summary
This summary is machine-generated.

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

Comparison of Common Plasma Proteomics Workflows Reveals Distinct Preanalytical Biases by Sample Preparation and Nanoparticle Enrichment.

Journal of proteome research·2026
Same author

Alpibectir-Ethionamide combination (AlpE) for the treatment of tuberculosis.

Nature communications·2026
Same author

Benchmarking EGF signaling pathway inference using phosphoproteomics and kinase-substrate interactions.

Nature communications·2026
Same author

High-throughput peptide-centric local stability assay extends protein-ligand identification to membrane proteins, tissues and bacteria.

Nature structural & molecular biology·2025
Same author

In silico biological discovery with large perturbation models.

Nature computational science·2025
Same author

Human gut bacteria bioaccumulate per- and polyfluoroalkyl substances.

Nature microbiology·2025
Same journal

Strain-Level Food Surveillance of <i>Escherichia coli</i> Using a Specific-Nonspecific Hybrid Sensor Array Strategy.

Analytical chemistry·2026
Same journal

A Field-Portable Fe(IV)-Mediated Competitive Quenching Chemiluminescence Platform with a Synchronous Y-Shaped Flow-through Cell for Broad-Spectrum Quantification of Volatile Phenols.

Analytical chemistry·2026
Same journal

Single-Molecule Characterization of CRISPR-Cas12a for Amplification-Free Genetic Testing.

Analytical chemistry·2026
Same journal

Integrated Acoustofluidic Manipulation and Oscillation-Stabilized Magnetic Relaxation Biosensing for <i>Salmonella</i> Detection.

Analytical chemistry·2026
Same journal

A Self-Powered Sensing Platform Based on the Janus Heterostructure for Machine Learning-Assisted Dual-Mode Detection of 17β-Estradiol.

Analytical chemistry·2026
Same journal

Large Language Model-Generated Dietary Metabolite Biomarker Database Drives Deep Annotation of the Human Diet Metabolome.

Analytical chemistry·2026
See all related articles

New isobaric mass tag proteomics methods increase sample multiplexing by using subtle mass differences. This allows for more comprehensive chemoproteomic kinase assays with greater compound concentration ranges in a single experiment.

Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Chemical Biology

Background:

  • Isobaric mass tag-based quantitative proteomics, including TMT (Tandem Mass Tag) and iTRAQ, uses reporter ions for relative quantification.
  • Multiplexing capacity in these methods is limited by the number of unique reporter ions generated through stable isotope incorporation.
  • Current TMT assays typically support 6-plex or 10-plex experiments.

Purpose of the Study:

  • To enhance multiplexing capabilities in isobaric mass tag quantitative proteomics.
  • To leverage high-resolution mass spectrometry and minor mass differences in reporter ions for increased sample capacity.
  • To improve the efficiency and scope of chemoproteomic kinase assays.

Main Methods:

  • Utilized the 6 mDa mass difference between (15)N- and (13)C-containing reporter fragments from TMT variants (TMT127L/H and TMT129L/H).

More Related Videos

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

Related Experiment Videos

Last Updated: May 19, 2026

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
16:10

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

  • Employed high-resolution mass spectrometry (Orbitrap Elite) with a 96 ms transient for baseline resolution of closely spaced reporter ions.
  • Applied the enhanced multiplexing strategy to chemoproteomic kinase assays.

    • Main Results:

    • Achieved baseline resolution of reporter ions differing by only 6 mDa, enabling higher multiplexing rates.
    • Demonstrated comparable dynamic range, precision, and accuracy to conventional 1 Da spaced reporter ions.
    • Enabled determination of inhibitor potencies across a wider range of compound concentrations in a single experiment.

    Conclusions:

    • The developed method significantly increases multiplexing capacity in isobaric mass tag proteomics.
    • This advancement allows for more comprehensive and efficient chemoproteomic profiling.
    • The strategy offers a powerful tool for drug discovery and chemical biology research.