iTRAQ Analysis
iTRAQ analysis is a high-throughput quantitative method widely used in proteomics research. By incorporating isotopic labels at peptide termini, this technique allows for the simultaneous relative and absolute quantification of proteins across multiple biological samples. The core principle of iTRAQ lies in its use of isotopic tags, which ensure identical mass signals for labeled samples in the mass spectrum. During collision-induced dissociation (CID), these tags release reporter ions with distinct masses, enabling quantitative comparisons across samples. This approach has found extensive applications in genomics and proteomics, particularly in disease biomarker discovery, drug target validation, and studies on protein expression dynamics in various biological processes.
In medical research, iTRAQ has been employed to investigate proteomic changes associated with diseases. By comparing healthy and diseased samples, researchers can identify proteins implicated in disease progression, providing essential biomarkers for diagnosis and treatment. In drug discovery, iTRAQ analysis facilitates the identification of drug-target proteins and elucidates the mechanisms underlying drug effects in cells or tissues. Additionally, iTRAQ plays a significant role in agricultural science, enabling the analysis of protein expression changes in plants under different environmental conditions to improve crop resilience and yield. In environmental science, iTRAQ is used to study the impact of pollutants on protein expression in organisms, providing critical insights for environmental protection. Overall, iTRAQ analysis offers a powerful tool for advancing research across various disciplines in life sciences.
The laboratory workflow for iTRAQ analysis involves several key steps. Initially, proteins are extracted and enzymatically digested from biological samples, followed by labeling peptides with iTRAQ reagents. The labeled peptides are then separated and analyzed using liquid chromatography-mass spectrometry (LC-MS). During MS analysis, iTRAQ-labeled peptides produce specific reporter ions upon CID, which serve as markers for quantification. Finally, researchers process the mass spectrometry data using specialized software to obtain accurate relative and absolute quantification of proteins. Successful application of iTRAQ requires well-designed experiments, precise sample preparation, and thorough data analysis. Experimental design should account for biological variability, include appropriate controls, and ensure sufficient replicates. Rigorous quality control during sample handling is critical to ensure accurate labeling and high sensitivity. Data analysis should utilize robust statistical methods and specialized software tools to accurately interpret the results.
The advantages of iTRAQ analysis include its high throughput, multiplex quantification capability, and exceptional sensitivity. Unlike traditional protein quantification methods that handle a limited number of samples, iTRAQ can simultaneously analyze multiple samples, significantly improving experimental efficiency. Moreover, it allows for the detection of low-abundance proteins, enhancing the characterization of complex proteomes in biological samples.
MtoZ Biolabs has a wealth of expertise in tag-based protein quantification methods, including iTRAQ, TMT, and SILAC. We are dedicated to delivering high-quality proteomics research services. Partnering with us provides clients with accurate quantitative analysis results, propelling their research projects forward. Whether for academic studies or clinical applications, we offer tailored solutions to meet diverse research requirements. Our comprehensive services, from sample preparation and experimental execution to data analysis, ensure reliable and efficient outcomes for all proteomics studies.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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