For cell samples that can be subcultured, we can take the SILAC method to complete the process of labeling the cell proteome, but many other research systems, such as tissue samples and cells that cannot be subcultured, require other labeling methods to complete quantitative proteomics analysis. Quantitative methods based on in vitro labeling such as iTRAQ have emerged.
iTRAQ refers to isobaric tags for relative and absolute quantitation technology. It is a new in vitro isotope labeling technology launched by AB SCIEX. This technology uses a variety of isotope reagents to label the N-terminal or lysine side chain groups of protein peptides. Mass spectrometer tandem analysis can simultaneously compare the differences in protein expression levels between multiple samples. It’s a high-throughput screening technology commonly used in quantitative proteomics recently.
iTRAQ reagent is composed of three parts: a reporter group, a balance group and a reactive group. The reactive groups form 2-8 isotopic labels of equal mass. iTRAQ labels the digested peptides, which can react with the N-terminal and primary amine groups of the side chain of the amino acid. In the primary spectrum, the same peptides from different samples show the same mass-to-charge ratio after labeling. In the secondary spectrum, the bond between the reporter group, the balance group and the reactive group is broken, and the quantitative information of the same peptide between samples can be obtained according to the abundance of the reporter ion, and then the quantitative information of the protein can be obtained through software processing.
Fig.1 Diagram of the iTRAQ workflow. (Romero R, et al. 2010.)
iTRAQ technology can accurately quantify and identify all proteins expressed in a genome or all proteins in a complex mixed system, search for differentially expressed proteins, and reveal their cellular physiological and pathological functions. iTRAQ technology is often used in the medical and health field. It can be used for large-scale experimental analysis, screening and verification of disease biomarkers, exploring protein expression regulation mechanisms during disease occurrence, and efficacy evaluation.
Creative Biogene’s iTRAQ quantitative proteomics service provides results including,
Our bioinformatics analysis covers a wide range including data quality control, protein identification analysis, protein quantitative analysis, protein pathway analysis, protein GO analysis and enrichment analysis, etc.
Cells, tissues, urine, whole blood, serum, plasma, total protein, etc.
In general, our turnaround time is 2-5 weeks depending on the size of your project.
Creative Biogene has excellent microbiology experts, well-established proteome technology platforms, and professional bioinformatics analysis to provide you with a one-stop service in iTRAQ quantitative proteomics. Our services include protein extraction, enzymatic hydrolysis, HPLC separation, mass spectrometric detection and bioinformatics analysis.
If you are interested in our services, please contact us for more details.