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Related Experiment Video

Updated: Jun 29, 2026

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

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

Published on: November 15, 2017

Improved proteome coverage by using iTRAQ labelling and peptide OFFGEL fractionation.

Emilie Ernoult¹, Erick Gamelin, Catherine Guette

¹Centre INSERM Régional de Recherche sur le Cancer U, Angers, France. emi.ernoult@wanadoo.fr

Proteome Science

|October 15, 2008

Summary

This study introduces a new shotgun proteomics method using isobaric tag for relative and absolute quantitation (iTRAQ) labeling and OFFGEL electrophoresis. This approach improves the identification of low-concentration proteins in complex biological samples.

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Last Updated: Jun 29, 2026

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

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Comprehensive Workflow of Mass Spectrometry-based Shotgun Proteomics of Tissue Samples

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Published on: November 13, 2021

Area of Science:

Proteomics
Mass Spectrometry
Biochemistry

Background:

Investigating complex protein mixtures from tissues is challenging due to wide dynamic concentration ranges.
Advanced mass spectrometry and fractionation methods are crucial for proteomic analysis.

Purpose of the Study:

To present a novel shotgun proteomics method combining iTRAQ labeling and OFFGEL electrophoresis.
To enhance the identification of low-concentration proteins in complex samples.

Main Methods:

Peptide fractionation using OFFGEL electrophoresis.
Isobaric tag for relative and absolute quantitation (iTRAQ) labeling of peptides.
Shotgun proteomics approach.

Main Results:

iTRAQ labeling enhances MALDI ionization efficiency.
OFFGEL fractionation adds isoelectric point (pI) as a validation criterion for protein identification.
iTRAQ-labeled samples enabled identification of lower-concentrated proteins compared to unlabeled samples.

Conclusions:

The combination of iTRAQ labeling and OFFGEL fractionation significantly increases proteome coverage for complex samples.
This method effectively supports the identification of low-concentration proteins.