Tandem Mass Tag (TMT)
Tandem Mass Tag (TMT) is a widely-used chemical labeling method in proteomics research, which leverages isotopic tags of varying masses to label proteins across different samples. These tags consist of three components: a reporter group, a balance group, and a reactive group. The reactive group specifically binds to amino groups on proteins, covalently attaching the tag. In tandem mass spectrometry, peptides labeled with Tandem Mass Tag (TMT) from different samples show identical mass in the primary spectrum. However, in the secondary spectrum, the tags cleave to produce distinct reporter ions. The detection and quantification of these reporter ions enable the relative quantification of identical proteins in different samples. Tandem Mass Tag (TMT) facilitates precise relative quantification of multiple samples in a single experiment by incorporating isotopic labels into protein samples. Additionally, TMT's design-which includes a reporter group generating detectable signals and a balance group minimizing mass differences-provides enhanced accuracy and sensitivity.
Tandem Mass Tag (TMT) technology can label up to 11 or 16 samples simultaneously within a single experiment and can be chemically modified to ensure precise distinction of tags during mass spectrometry. This multiplexing capability allows Tandem Mass Tag (TMT) to excel in the analysis of complex samples, such as comparing tumor tissues with normal tissues in cancer research to identify differentially expressed proteins. Tandem Mass Tag (TMT) also holds significant potential in biomarker discovery, drug mechanism investigation, and disease diagnosis and prognosis.
Tandem Mass Tag (TMT) Technology Workflow
1. Sample Preparation and Protein Extraction
Proteins must be extracted from samples prior to Tandem Mass Tag (TMT) labeling, ensuring protein integrity and consistent concentration.
2. Protein Digestion and Labeling
Proteins are digested with trypsin into peptides, which are then labeled with Tandem Mass Tag (TMT) reagents. Reaction conditions must be optimized for efficient and precise labeling.
3. High-Performance Liquid Chromatography (HPLC) Separation
The peptide mixture undergoes separation via HPLC, reducing background interference and improving mass spectrometry accuracy.
4. Mass Spectrometry Analysis
Peptide samples are analyzed using mass spectrometry, where Tandem Mass Tag (TMT)-generated peaks facilitate quantitative analysis.
5. Data Analysis and Interpretation
Bioinformatics tools are employed to analyze mass spectrometry data, yielding relative quantification and functional annotations between samples.
Considerations and Common Issues
1. Labeling Efficiency
Variations in labeling efficiency across samples should be accounted for to prevent inaccuracies in data.
2. Sample Complexity
Complex samples may challenge separation and analysis, requiring optimized pretreatment and separation strategies.
3. Data Processing
Processing Tandem Mass Tag (TMT) data involves specialized software to accurately interpret information from multiple samples.
Tandem Mass Tag (TMT) allows for high-throughput analysis of multiple samples in a single experiment, ensuring high sensitivity and accuracy. This technology is versatile and applicable to a range of biological samples, such as cells, tissues, and fluids. MtoZ Biolabs offers high-resolution mass spectrometry platforms for premium Tandem Mass Tag (TMT) analysis services. Our expert team specializes in complex sample handling, guaranteeing precise and reliable quantitative results. MtoZ Biolabs is dedicated to providing tailored solutions for both basic research and applied development needs.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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