Mitochondrial TMT & iTRAQ & MultiNotch Analysis

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Isobaric tags for relative and absolute quantitation (iTRAQ) technique and Tandem mass tags (TMT) technique are in vitro labeling quantitative techniques for peptides developed by American AB Sciex and Thermo Fisher, respectively. These two techniques use multiple (2-10) stable isotope tags and specifically label the amino groups of peptides for tandem mass spectrometry analysis, which can be used to compare the relative content of proteins in as many as 10 different samples at the same time. It can be used to study the differences in protein expression levels in tissue samples under different pathological conditions or different developmental stages.

For cell samples that can be passaged, we can use SILAC to complete the labeling process of cell proteome, but many other research systems, such as tissue samples and cells that cannot be subcultured, need other labeling methods to complete quantitative proteomic analysis. TMT, iTRAQ, and other quantitative methods based on in vitro labeling came into being.

ITRAQ is an isotope labeling reagent. ITRAQ reagent consists of three parts: reporter group, mass balance group, and peptide reaction group, forming four or eight ectopic tags with equal relative molecular weight (145 or 305).

Figure 1. iTRAQ reagent structures of 4 isotope tags (A) and 8 isotope tags (B).

The same peptide from different samples had the same mass number after labeled with TMT/ iTRAQ reagent and showed the same mass peak in the first-order mass spectrometry (MS). When the mass peak is selected for fragmentation, in the secondary mass spectrometry (MS/MS), different reporter groups are released, and the signal intensity of their respective mass peaks represents the expression level of the peptide and its corresponding protein from different samples. TMT/ iTRAQ quantitative proteomics research has been widely used because of its high accuracy and adaptability to a wide variety of samples.

Figure 2. TMT technical process.

Based on traditional TMT/ iTRAQ analysis technology, Creative Biogene provides TMT/ iTRAQ/ MultiNotch MS analysis technology and uses tertiary fragment report ions for quantitative analysis, which can further improve the accuracy of TMT/ iTRAQ quantitative proteomic analysis and quantitative accuracy.

Creative Biogene uses Thermo Fisher's Q ExactiveHF mass spectrometry platform, Orbitrap Fusion Lumos mass spectrometry platform, and Nano-LC, to launch quantitative proteome analysis service technology based on TMT/ iTRAQ/ MultiNotch MS. You only need to tell us the purpose of your experiment and send us your protein. We will be responsible for all the follow-up matters of the project, including protease digestion, peptide pricing, peptide separation, mass spectrometry analysis, mass spectrometry raw data analysis, bioinformatics analysis.

Work Flow

Creative Biogene will provide you with a detailed technical report when the project is delivered, including experimental steps, relevant MS parameters, details of identified amino acids, MS images (high resolution), and detailed raw data. Creative Biogene always provides you with customized services for mitochondrial research, and our one-stop research and analysis platform can meet all your research needs.

Technical characteristics

• High-throughput quantitative protein analysis: it can analyze 8-10 samples at the same time; it is suitable for large-scale experimental analysis of a large number of clinical samples and multi-control groups, and new biomarkers are found.
• In vitro labeling, suitable for the analysis of tissue, cell, blood, and other samples.
• Mass spectrometry analysis has high sensitivity, can detect low abundance proteins, and can compare the relative abundance of proteins in a wide range.
• It can be combined with the methods of enriching modified peptides such as phosphorylation.

Our Advantages

• Rich experience in mitochondrial quantitative proteomics research
• Outstanding research team
• Unique integrated service for mitochondrial research
• Reliable data and results
• Rapid turnaround and cost-effective

Creative Biogene has the most professional team to serve customers around the world. If you have any questions about the content of this service, please feel free to contact us. We look forward to your contact.

Reference

1. Jiang P.J.,et al. “ Quantitative proteomics analysis of differentially expressed proteins in ruptured and unruptured cerebral aneurysms by iTRAQ.” Journal of Proteomics, 2018, 182: 45-52.

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