Introduction:

iTRAQ (4 plex or 8 plex) and TMT (2 plex through 16 plex) are chemical labeling methods that enable high through-put and internally controlled quantifications in proteomics studies. This is a very popular service in Complete Omics which enables researchers to directly compare the difference of proteomes in their treated vs untreated, diseased vs normal, over-expressed vs control, gene silenced vs control, knockout vs wild type, mutant vs wild type groups, etc. ​

Every sample group in the study is normalized to start from the equal amount of total protein/peptide. Protein digest of each sample is chemically labeled with a sample specific tag. Tagged peptides from different samples are pooled together with equal total individual amount and the complete catalogue of proteins in the pooled sample is resolved by Mass Spectrometry analysis and each protein identified in the study will contain 4 or 8 (for iTRAQ) and 2 through 16 (for TMT) intensity values representing this protein’s abundance in different samples. The result shows not only the identification of peptides/proteins, but also the absolute/relative abundance of the peptides/proteins in each sample.

Your samples (several test groups and control group) will be digested with Trypsin and peptides are chemically labeled by either iTRAQ or TMT labeling method, and the labeled peptides are pooled and ion-exchanged through Strong Cation Exchange (SCX) HPLC cleaning. To improve the sensitivity, we suggest the basic Reverse Phase LC (bRPLC) to further fractionate your cleaned sample into 96 fractions and then pooled to 24 fractions to maximize the coverage and depth of your analysis. 24 Mass Spectrometry Analysis runs will be performed by the world’s most advanced Orbitrap LC-MS/MS system. 

Data analysis, including data validation, visualization and quantification, are performed with commercial softwares and Complete Omics’ unique R packages and scripts. Report will be sent to you in Excel format as well as a summary in PDF format. We will also provide you any details you need for your papers’ MATERIALS AND METHODS section. We will make sure you understand your result and help you with your paper writing with free follow-up services.

Sample types we accept:

1, 2D gel spots, SDS-PAGE bands

2, Cell Lysates and Tissue Lysates

3, Biofluids, such as plasma*, serum*, saliva, tear, etc.

*We provide High Abundance Protein Depletion Service to significantly (100-500 folds) increase the depth of your biofluid proteomics analysis by removing top abundant proteins from your samples. Read more for details​.

4, FFPE slide/ FFPE extract 5, Customized sample types (please contact us to discuss)

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