iTRAQ-Based Proteomics Analysis

Creative Proteomics has an impressive track record in quantitative proteomics analysis. We provide advanced high-throughput iTRAQ-based proteomics analysis service to provide technical support to those engaged in protein research.

iTRAQ (isobaric tags for relative and absolute quantitation) uses multiple isobaric tags for relative and absolute quantitation of proteins. This is a patented technology for proteome quantification developed by AB SCIEX in the United States. It can be applied to protein identification and quantification in a variety of samples such as bacteria, yeast, human tissue, cells, and body fluids, and can simultaneously compare the expression levels between 2 to 8 protein samples. As a new quantitative proteomics research method, this technology exhibits high throughput, good reproducibility, and high sensitivity, and has been widely used to study the pathogenesis of many infectious pathogens.

Service Content of iTRAQ-Based Proteomics Analysis

In the iTRAQ-based proteomics analysis service, Creative Proteomics utilizes isobaric reagents to label primary amines of peptides and proteins. The role of the balancing group in iTRAQ reagents is to make the labeled peptide in each sample isobaric and to facilitate quantification by analyzing the reporter group generated upon fragmentation in the mass spectrometer. This greatly facilitates peptide identification due to the higher intensities of the precursor and fragment ions. Identify and quantify peptides in the same experiment using multidimensional LC and MS/MS by comparing four MS/MS reporter ions (range 114 to 117 Da) or eight MS/MS reporter ions (range 113-119 and 121 Da) peak area and the resulting peak ratio to obtain mass spectrometry results. In this service, we can provide sample preparation services such as cell lysis, trypsin hydrolysis of proteins to produce tryptic peptides, etc. Bioinformatics analysis services such as functional annotation and cluster analysis can also be provided.

Mass Spectrometry Facilities and Projects

• AB SCIEX Triple Quad™ 6500 Mass Spectrometer
• AB SCIEX QTRAP 5500 Ion Trap Mass Spectrometer
• AB Sciex API 3000 ™ LC/MS/MS Triple Quadrupole Tandem Mass Spectrometer

Requirements for Samples

• Samples must be prepared in buffers that do not contain primary amines.
• The recommended sample size is 25 µg (8 plex) to 100 µg (4 plex).
• Quantify at the MS/MS spectral level, typically 400 or more identified proteins from the crude cell or tissue extracts.
• Please provide the specific concentration, volume, preparation time, and source of each sample. And inform the sample information as well as the control and experimental samples (if there is a group, describe the group information in detail.)

Deliverables

• Experimental steps
• Relevant mass spectrometry parameters
• Details of the identified phosphorylation sites
• Mass spectrometry images
• Raw data

Our Advantages

• The labeling efficiency is high, and almost all proteins in the sample can be labeled.
• Mass spectrometry has high detection sensitivity and can detect low-abundance proteins.
• Accurate quantification of multiple protein mixtures or recombinant protein mixtures is possible.

Service Process

With advanced mass spectrometry facilities and an experienced team, Creative Proteomics can provide researchers with professional protein quantitative analysis services. If you are interested in our iTRAQ-based proteomics analysis service, please contact us directly for details and quotation.