iTRAQ and TMT Based Quantitative Proteomics by LC-MS/MS

Proteomics technologies that rely on mass spectrometry remain a powerful technique that can be used to characterize biological and clinical samples at the protein level. The Isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and Tandem Mass Tag (TMT) are validated chemical labels used for gel-free quantitative proteomics by mass spectrometry. The iTRAQ & TMT technologies permit the simultaneous identification and relative quantitation of proteins in 2 to 8 different samples in a single multiplexed experiment.

  • Using this powerful gel-free proteomics approach it is possible to a) identify 100s to 1000s of proteins in 2 to 8 samples in a single LC-MS/MS run, and b) simultaneously quantify the relative concentrations of the identified proteins.

The Advantages of this proteomics approach include:
1) Increased throughput.
2) The ability to identify and simultaneous quantify proteins in one single run.
3) 2-8 samples can be labeled and multiplexed into a single sample prior to a single LC-MS/MS run.
4) A large dynamic range which allows the identification of high and low abundant proteins of different diversities.

Our iTRAQ or TMT service will include the following:
1) Sample preparation
2) Sample digestion with trypsin or a suitable enzyme.
3) 2 - 8 Plex iTRAQ or TMT labeling in which we use from 5 to 100µg per sample.
4) Pooling of labeled samples
5) Fractionation of samples by strong cation exchange (SCX) chromatography to increase sensitivity.
6) Tandem mass spectrometry analysis using high performance nano-LC/MS/MS.
7) Database search and Identified full protein annotation.
8) Report generation.

 

 


Important Info

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