iTRAQ Labeling
iTRAQ labeling (Isobaric Tags for Relative and Absolute Quantitation) employs isotopic tags to facilitate the simultaneous quantitation of up to eight proteomic samples. By chemically tagging peptides, this method allows for precise quantification of protein abundance using mass spectrometry. iTRAQ's high quantification efficiency makes it especially valuable in biomedical research, including the investigation of complex biological systems and disease mechanisms. Its applications span the discovery of disease biomarkers, validation of drug targets, and examination of dynamic biological processes. Through iTRAQ labeling, differential protein expression can be elucidated under diverse physiological or pathological conditions, enhancing our understanding of disease pathogenesis. Moreover, iTRAQ showcases exceptional performance in the analysis of clinical samples, assisting researchers in identifying proteins with differential expression across extensive datasets, thus supporting personalized and precision medicine.
Process of iTRAQ Labeling Technology
1. Protein Extraction and Enzymatic Digestion
Extracted proteins are enzymatically digested, commonly with trypsin, to generate peptides. Complete digestion is crucial for comprehensive peptide profiling.
2. iTRAQ Reagent Labeling
Peptides are reacted with iTRAQ reagents, each sample receiving distinct isotopic tags. This requires stringent reaction condition control to ensure effective and uniform labeling.
3. Peptide Separation and Purification
Labeled peptides are fractionated and purified using liquid chromatography to decrease sample complexity and improve mass spectrometric accuracy.
4. Mass Spectrometry Analysis
Post-separation, peptides undergo mass spectrometric analysis to generate spectra. High-resolution mass spectrometry is essential for distinguishing isotopically labeled peptides.
5. Data Analysis and Interpretation
Mass spectrometric data are analyzed using specialized software to determine protein abundance quantitatively. The reliability of research outcomes hinges on the application of validated data processing algorithms.
Considerations and Challenges
Standardized sample preparation is vital to minimize bias. High labeling efficiency and reproducibility are critical for methodological success. The complexity of mass spectrometry data necessitates skilled analysis.
MtoZ Biolabs provides bespoke iTRAQ labeling services, tailored to client specifications, offering comprehensive solutions from sample preparation to data interpretation. Collaborate with MtoZ Biolabs for cutting-edge research support.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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