Can Quantitative Proteomics TMT/iTRAQ Identify the Presence of Proteins?
Quantitative Proteomics is a technique in biology mainly used for quantitatively comparing the relative or absolute abundance of proteins under different conditions. Among them, TMT (Tandem Mass Tag) and iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) are the most commonly used labeling quantitative techniques. They achieve protein quantification by adding tags to proteins or peptides, which produce specific reporter ions in mass spectrometry. So, can TMT/iTRAQ identify the presence or absence of proteins?
1. Basic Principles of TMT/iTRAQ
TMT/iTRAQ is an isotope labeling-based quantitative proteomics technology. Its basic steps include protein extraction, digestion, labeling, mixing, chromatographic separation, and mass spectrometry analysis. TMT/iTRAQ tags are used to label the N-terminus of peptides and the ε-NH2 of lysine, consisting of a balance group and a reporter group. The balance group mass remains constant, while the reporter group mass varies, producing reporter ions of different masses, enabling simultaneous quantification of multiple samples.
2. Can TMT/iTRAQ Identify the Presence or Absence of Proteins?
TMT/iTRAQ technology can quantitatively identify the differences in protein abundance across different samples, reflecting the presence or absence and increase or decrease of proteins. However, it should be noted that TMT/iTRAQ is a relative quantification technique; it measures the relative abundance of proteins between different samples and cannot directly reflect the absolute abundance of proteins in a sample. Therefore, TMT/iTRAQ can be used to identify the presence or absence of specific proteins under specific conditions, but determining the absolute content of proteins requires combining with other methods, such as full standardization techniques.
3. Applications of TMT/iTRAQ
1. Revealing Dynamic Changes in Biological Processes: TMT/iTRAQ can quantitatively obtain changes in protein abundance at different time points, under different environments, or during different biological processes, thereby revealing the dynamic changes in biological processes.
2. Discovery of Disease Biomarkers: TMT/iTRAQ can be used to compare the differences in protein expression between disease and healthy groups, aiding in the discovery of disease biomarkers.
3. Research on Drug Mechanisms: By comparing the differences in protein expression between drug-treated and control groups, the mechanism of drug action can be revealed.
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Quantitative Proteomic Analysis
Label-based Protein Quantification Technologies - iTRAQ, TMT, SILAC
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