iTRAQ/TMT

iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) is a mass spectrometry-based technique for protein quantification. This method uses chemical reagents to label peptides in different samples with isotopic tags, allowing for the relative and absolute quantification of proteins. iTRAQ can compare protein expression levels in up to eight different samples simultaneously. This technology is particularly useful for analyzing proteins in complex samples, such as cell lysates, tissue extracts, and serum. Due to its high throughput and multiplexing capabilities, iTRAQ is widely used in proteomics research.

 

TMT (Tandem Mass Tag) is an isotope labeling technique used in mass spectrometry for the relative quantification of proteins or peptides. TMT involves the introduction of chemical tags into samples, enabling the identification and quantification of the same proteins or peptides across different samples using mass spectrometry. Each TMT tag consists of a reporter ion, a balance group, and a reactive group. This technology is suitable for various proteins or peptides, especially in complex biological samples, large-scale proteomics studies, and biomarker discovery.

 

iTRAQ Technology Analysis Workflow

1. Sample Preparation: Extract proteins and perform enzymatic digestion.

 

2. Tag Labeling: Label peptides with iTRAQ reagents.

 

3. Sample Mixing: Mix labeled peptides from different samples.

 

4. LC-MS/MS Analysis: Separate and detect peptides using liquid chromatography-tandem mass spectrometry.

 

5. Data Processing and Analysis: Use software to analyze mass spectrometry data, quantifying peptides and proteins across samples.

 

TMT Technology Analysis Workflow

1. Sample Preparation: Extract proteins and enzymatically digest to generate peptides.

 

2. Tag Labeling: Label peptides in different samples with TMT reagents.

 

3. Sample Mixing: Combine labeled samples.

 

4. Liquid Chromatography Separation: Separate mixed samples using a liquid chromatography system.

 

5. Mass Spectrometry Analysis: Perform analysis using an LC-MS/MS system, collecting reporter ion data from TMT tags.

 

6. Data Processing: Utilize specialized software for data processing and quantification.

 



Griss, J. et al. J Proteome Res. 2019.

Figure 1. Scheme of the Entire Workflow for iTRAQ and TMT

 

Applications

1. Disease Research: Compare disease group and control group samples to identify potential disease-related proteins.

 

2. Drug Development: Study protein expression changes before and after drug treatment to evaluate drug mechanisms.

 

3. Functional Studies: Reveal protein expression changes under specific conditions to study protein functions.

 

4. Biomarker Research: Identify and quantify disease-related biomarkers using iTRAQ and TMT technologies.

 

5. Multiplex Sample Quantification: Simultaneously quantify proteins or peptides in multiple samples, improving analytical efficiency.

 

6. High Throughput Analysis: Suitable for large-scale proteomics research, capable of quantifying hundreds to thousands of proteins.

 

Service Advantages

1. High Throughput: Simultaneous comparison of multiple samples increases experimental efficiency.

 

2. High Sensitivity: Detection of low-abundance proteins in complex samples.

 

3. High Accuracy: Improved quantification accuracy and repeatability through internal standard correction.

 

4. Multiplex Labeling: Compare up to eight samples simultaneously, enhancing data richness and comprehensiveness.

 

5. Precise Quantification: Enhanced accuracy and reproducibility using internal and external standards.

 

6. Good Data Consistency: Reduced inter-experimental variability as samples are analyzed in the same mass spectrometry run.