Protein Sequencing for TMT

Tandem Mass Tag (TMT) protein sequencing is an advanced mass spectrometry technique used to quantitatively compare the expression levels of proteins in different samples. TMT tags are a set of chemical markers that can covalently bind to the amino terminus and lysine side chains of proteins or peptides. Through mass spectrometry analysis, these tags allow the simultaneous quantitative analysis of protein expression differences in up to 16 different samples. TMT protein sequencing technology is particularly suitable for proteomics research in complex biological samples, including disease mechanism research, drug mechanism exploration, and the discovery of biological markers.

 

Key Steps of TMT Protein Sequencing

1. Sample Preparation

Proteins are extracted from the samples to be analyzed and digested with an enzyme (usually trypsin) to produce a peptide mixture.

 

2. Labeling

Different TMT tags are added to the peptides of different samples respectively. Each TMT tag has a unique mass that allows the labeled peptides to be distinguished in mass spectrometry analysis.

 

3. Mixing

All labeled samples are mixed together so that multiple samples can be processed simultaneously in the same mass spectrometry analysis.

 

4. Liquid Chromatography Separation (LC)

The mixed peptide samples are separated by liquid chromatography to reduce sample complexity and improve the resolution and sensitivity of mass spectrometry analysis.

 

5. Mass Spectrometry Analysis

Analysis is performed by a mass spectrometer to measure the mass and fragment patterns of the peptides. TMT tags allow the relative levels of the same protein in different samples to be quantitatively compared by detecting the relative intensities of the labeled peptides.

 

6. Data Analysis

Specialized software is used to process and analyze the mass spectrometry data, identify proteins and quantitatively compare their expression differences in different samples.

 

Advantages

1. High-Throughput

Multiple samples can be analyzed simultaneously, greatly improving experiment efficiency.

 

2. High Sensitivity

TMT tags enhance the ability to detect low-abundance proteins.

 

3. High Accuracy

The accuracy and reproducibility of quantitative analysis are high.

 

Applications

TMT protein sequencing is widely used in proteomics research, especially in the identification of disease biomarkers, the exploration of drug targets, and the study of basic biological processes. By comparing protein expression differences under healthy and diseased states, at different time points, or under different treatment conditions, researchers can gain important information about biological mechanisms.