iTRAQ (Isobaric Tag for Relative and Absolute Quantification) andTMT(Tandem Mass Tags) technology are in vitro peptide labeling quantification technologies developed by AB Sciex and Thermo Fisher in the United States. These technologies use multiple (2-10) stable isotope tags to specifically label the amino groups of peptides for tandem mass spectrometry analysis, allowing for the simultaneous comparison of the relative protein content in up to 10 different samples. They are used to study differences in protein expression levels in tissue samples under different pathological conditions or at various developmental stages.
For cell samples that can be passaged, we can useSILACmethod to complete the labeling process of the cellular proteome. However, for many other research systems, such as tissue samples and cells that cannot be passaged, other labeling methods are needed to complete quantitative proteomics analysis. TMT, iTRAQ, and other in vitro labeling-based quantification methods have emerged.
The iTRAQ tag consists of three parts, as shown below:
1. Reporter group: Indicates the abundance level of protein samples.
2. Balance group: Balances the mass difference of the reporter group to ensure that the isobaric tags have the same weight, ensuring that the labeled peptide segments have the same m/z.
3. Amine-specific reactive group: Capable of covalently bonding with the N-terminal and lysine side chain amino groups of peptides, thereby tagging the peptide segments.
iTRAQ and TMT tag structural diagram
The same peptide segments from different samples, after being labeled with TMT/iTRAQ reagents, have the same mass and appear as a single mass spectrometry peak in MS1. When this peak is selected for fragmentation, different reporter groups are released in MS2. The signal intensity of each mass spectrometry peak represents the expression levels of the peptide and its corresponding protein from different samples. TMT/iTRAQ quantitative proteomics is widely used due to its high accuracy and adaptability to various sample types. Based on traditional TMT/iTRAQ analysis technology, BiotechPacker provides TMT/iTRAQ/MultiNotch MS analysis technology, using tertiary fragment reporter ions for quantification, further improving the accuracy of TMT/iTRAQ quantitative proteomics analysis.
BiotechPacker uses Thermo Fisher's Q Exactive HF mass spectrometry platform, Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC, to offer a quantitative proteomics analysis service package based on TMT/iTRAQ/MultiNotch MS. You just need to tell us your experimental goals and send us your proteins, and we will handle all subsequent project matters, including protein digestion, peptide labeling, peptide separation, mass spectrometry analysis, raw data analysis, and bioinformatics analysis.
iTRAQ/TMT quantitative proteomics analysis technical features
• High-throughput quantitative protein analysis: Can analyze 10 samples simultaneously; suitable for large-scale experimental analysis with a large number of clinical samples and multiple control groups, discovering new biomarkers.
• In vitro labeling applicable to the analysis of various samples such as tissues, cells, and blood.
• High sensitivity of mass spectrometry analysis, capable of detecting low-abundance proteins and comparing protein relative abundance over a wide range.
• Can be combined with methods for enriching phosphorylated and other modified peptides to study post-translational modifications more accurately.
Note: For sample transport, please use an adequate amount of dry ice and choose a faster shipping method to reduce the possibility of sample degradation during transport.
Bilingual project report
In the technical report, BiotechPacker will provide you with a detailed bilingual (Chinese and English) technical report, including:
1. Experimental steps (in Chinese and English)
2. Relevant mass spectrometry parameters (in Chinese and English)
3. Mass spectrometry images
4. Raw data
5. Protein differential level analysis
6. Bioinformatics analysis
