iTRAQ Quantification Technology: Principles, Advantages, and Applications
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Labeling process: The iTRAQ reagent comprises a reporter group, a balancer group, and a peptide-reactive group. Through the amine-reactive moiety, the iTRAQ tag covalently binds to the N-terminus and lysine residues of peptides.
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Mass spectrometry detection: During MS/MS analysis, the labeled peptides undergo collision-induced dissociation (CID), generating reporter ions of distinct mass-to-charge ratios. Quantification is based on the signal intensities of these reporter ions, from which the relative protein abundance across samples can be inferred.
In proteomics research, the accurate and sensitive comparison of protein abundance across different samples is crucial for elucidating underlying biological mechanisms. iTRAQ (Isobaric Tags for Relative and Absolute Quantitation), a mass spectrometry-based multiplex quantification technique, has become a pivotal tool in life sciences due to its high throughput, precision, and wide applicability. This paper systematically introduces the principles, key advantages, and major applications of iTRAQ technology.
Basic Principles of iTRAQ Quantification Technology
iTRAQ is an isotope labeling technique in which peptides from different samples are chemically tagged so that they exhibit identical masses during the MS1 scan of mass spectrometry, yet release distinct reporter ions upon fragmentation, enabling the relative quantification of multiple samples.
Main Advantages of iTRAQ Quantification Technology
1. Simultaneous Quantification of Multiple Samples
iTRAQ enables the concurrent labeling and analysis of up to 8 or more sample groups, significantly enhancing experimental throughput. It is particularly well-suited for complex biological systems, including disease progression studies and time-course analyses.
2. High Quantification Accuracy and Sensitivity
Utilizing a standardized mass spectrometry workflow along with internal reference controls, iTRAQ allows for precise detection of low-abundance proteins and effectively minimizes technical variability.
3. Broad Applicability
iTRAQ is compatible with cellular samples, tissue specimens, and various biological fluids. When combined with high-resolution mass spectrometry platforms, it further improves both the depth of protein identification and the accuracy of quantification.
Applications of iTRAQ Quantification Technology
1. Disease Mechanism Research
iTRAQ quantification technology has been widely applied in studies of cancer, cardiovascular diseases, and neurological disorders. It enables the identification of differentially expressed proteins, thereby elucidating the molecular mechanisms underlying disease onset and progression.
2. Biomarker Discovery
By comparing protein expression profiles between patient and control samples, iTRAQ facilitates the discovery of early diagnostic biomarkers and therapeutic targets, offering foundational data to support precision medicine.
3. Mechanism Analysis of Drug Action
iTRAQ is employed to assess changes in protein expression before and after drug treatment, revealing the mechanisms of drug action as well as potential pathways of adverse effects.
4. Plant and Microbiome Research
iTRAQ is also extensively utilized in agricultural science and the study of environmental microbial communities, contributing to the advancement of fundamental biological research.
With the continuous advancement of proteomics, iTRAQ quantification technology—thanks to its multiplexing capabilities and high precision—has become a vital tool for addressing key questions in life sciences. In the future, as mass spectrometry and data analysis technologies continue to evolve, iTRAQ is expected to demonstrate significant application potential across an even broader spectrum of research areas. MtoZ Biolabs is dedicated to delivering high-quality quantitative proteomics analysis services, supporting your scientific journey through expertise and innovation.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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