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Updated: May 19, 2026
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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
An accurate proteomic quantification method: fluorescence labeling absolute quantification (FLAQ) using
Junyan Liu1, Yang Liu, Mingxia Gao
1Department of Chemistry & Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Proteomics
|August 14, 2012
Summary
A new method called fluorescent labeling absolute quantification (FLAQ) offers precise protein quantification using chromatography and mass spectrometry. This technique achieves high accuracy and sensitivity, enabling detailed proteomic analysis.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Accurate protein quantification is crucial for understanding biological processes.
- Existing methods often rely solely on mass spectrometry (MS), which can have limitations.
Purpose of the Study:
- To develop a facile and accurate method for absolute protein quantification.
- To combine chromatography-based quantification with MS for protein identification.
Main Methods:
- Developed the fluorescent labeling absolute quantification (FLAQ) method.
- Employed multidimensional liquid chromatography (MDLC) with laser-induced fluorescence (LIF) detection and tandem MS.
- Utilized 5-iodoacetamidofluorescein for labeling cysteine residues.
Main Results:
- Achieved high accuracy (RSD% < 1.58) and wide linearity (1-10^5) in quantification.
- Demonstrated a low limit of quantitation (0.34 amol) for model proteins.
- Successfully quantified proteins in a human liver proteome sample.
Conclusions:
- FLAQ is a robust method for absolute protein quantification.
- The method offers high sensitivity and accuracy, complementing MS identification.
- FLAQ has potential applications in various proteomic studies.
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