TMT Protein Quantitation Service

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Creative Proteomics' tandem mass tag (TMT) in vitro labeling-based mass spectrometry platform provides protein quantification services to help find differentially expressed proteins.

TMT are composed of three parts: molecular weight reporter, molecular weight normalization part and reaction group, forming two, six or ten kinds of ectopic tags with equal relative molecular mass. The TMT reagent can efficiently label the enzymatic peptides through the reactive group. In the first-order mass spectrum, the molecular weight normalization part makes the same peptide in different samples labeled with any one of the TMT reagents show the same mass-to-charge ratio. In tandem mass spectrometry, the cuttable arms can preferentially break to release molecular weight reporters. Each reporter has its own unique molecular weight, and can reflect the sample abundance of the labeled peptide in MS / MS analysis. The quantitative information of the same peptides between samples can be obtained based on the signal intensity value of the reporter ion, and then passed through the software Processed to get quantitative information on the protein.

The Workflow of TMT Protein Quantitation Service

The process for MS experiments using TMT are summarized as follows. protein extracts isolated from cells or tissues are reduced, alkylated, and then digested using protein lysate or equivalent. The sample is then labeled with TMTpro reagent before the sample is mixed, fractionated, and purified. Prior to data analysis, labeled samples were analyzed on a high-resolution Orbitrap LC-MS / MS mass spectrometer to identify peptides and quantify the relative abundance of the reported ions.

TMT Protein Quantitation Service

Technology Platform

Orbitrap LC-MS/MS Mass Spectrometer.

Advantages of TMT Protein Quantitation Service:

Increased sample throughput for parallel MS analysis for multiple samples, enabling the simultaneous quantitative analysis of up to 11 (TMT) or 16 (TMTpro) different samples from different samples from cells, tissues or biological fluids.
Improved multi-channel analysis capabilities reduce loss of quantitative data.
A variety of chemical groups can be used to specifically label different functional groups, and the labeled functional groups can be specifically enriched with anti-TMT solid phase resin.
Amine, cysteine, and carbonyl-reactive chemicals can use effective strategies to efficiently label target functional groups.
Optimized for high-resolution MS/MS platforms using SP2 or SPS MS3 quantitation mode, Proteome Discoverer software fully supports corresponding data analysis to improve multi-channel analysis capabilities and reduce loss of quantitative data.
With high throughput, one assay can simultaneously obtain the quantitative and quantitative information of thousands of proteins.
The TMT experiment cycle is short, which can reduce the entire experiment cycle and the experimental error between samples.
Compared with gel-separated protein quantification technology, TMT mass spectrometry technology can detect more types of protein molecules, such as low-abundance proteins, strong acid (basic) proteins, 10KD or 100KD protein molecules.
The fragmentation efficiency of the second mass spectrometry is more conducive to the analysis of protein interactions and expression patterns.

Sample Requirements

Plant leaf samples: wet weight≥1g.
Plants rich in impurities or samples with low protein content, such as plant roots, rhizomes, xylem, phloem tissue: dry weight≥5g.
The number of cell samples must reach 107 (It is recommended to use lysate for lysis before sending).
Fresh human, animal and plant tissue:≥100mg.
Serum/plasma≥500ul.
Urine≥25mL.
Body fluids such as saliva, amniotic fluid, cerebrospinal fluid≥5mL.

For other special samples, please contact us if you have any questions.

Application of TMT Protein Quantitative Technology

Screening for disease markers
Molecular mechanism research
Plant stress resistance research
Drug target research
Special function protein screening

Delivery

Complete proteomics experiment report (including specific experimental procedures), target protein candidate peptide secondary mass spectrum matching map, skyline analysis of target peptide TMT results, quantitative results and statistical analysis of target peptides and proteins.

Want to Know about Other Proteomics Quantitative Analysis Techniques?

References

Venable JD, Steckler C, Ou W, et al. (2015) Isotope-Coded Labeling for Accelerated Protein Interaction Profiling Using MS. Anal Chem,2015, 87:7540−7544.
Yu C, Huszagh AS, Viner R, et al. Developing a Multiplexed Quantitative Cross-linking Mass Spectrometry Platform for Comparative Structural Analysis of Protein Complexes. Anal Chem, 2016, 88(20):10301–10308.

* For Research Use Only. Not for use in diagnostic procedures.