iTRAQ Proteomics: Unveiling Molecular Mechanisms of Proteomics

Proteomics is a scientific field that studies the composition, structure, and function of all proteins in a biological body. In recent years, with the development of biotechnology, proteomics has played an increasingly important role in biomedical research. itraq proteomics is a commonly used protein quantification technology that reveals the molecular mechanisms of proteomics through its unique principles.

 

itraq (Isobaric Tags for Relative and Absolute Quantitation) proteomics is a mass spectrometry-based protein quantification technology. The basic principle is to use a chemical labeling method to label proteins in different samples with isotopic labels of the same mass. These isotopic labels produce specific mass peaks during mass spectrometry analysis. By measuring the intensity of these mass peaks, the abundance differences of proteins in different samples can be quantitatively analyzed.

 

Steps of Itraq Proteomics

The experimental process of itraq proteomics includes sample preparation, protein digestion, labeling, mixing, separation, and mass spectrometry analysis.

 

1. Sample Preparation

Firstly, proteins need to be extracted from different samples and dissolved in an appropriate buffer. The quality and purity of sample preparation are crucial to the results of subsequent experiments.

 

2. Protein Digestion

The protein sample is digested, and the commonly used enzyme is trypsin. After digestion, the proteins are cut into small peptides, which are easy for subsequent mass spectrometry analysis.

 

3. Labeling

The peptides in different samples are labeled with itraq reagents respectively. itraq reagents are chemical reagents containing isotopic labels, which can react with the amino groups in peptides to form labeled peptides.

 

4. Mixing

The labeled peptides are mixed together to form a composite sample. The mixing ratio can be adjusted according to the needs of the experimental design.

 

5. Separation

Technologies such as high-performance liquid chromatography (HPLC) are used to separate the peptides in the composite sample. The purpose of separation is to reduce complexity and make mass spectrometry analysis more accurate and reliable.

 

6. Mass Spectrometry Analysis

The separated peptides are analyzed by mass spectrometry, and the commonly used technology is liquid chromatography-tandem mass spectrometry (LC-MS/MS). Through mass spectrometry analysis, the mass peaks of peptides can be obtained and their intensity can be measured.

 

Applications

Itraq proteomics technology has a wide range of applications in biomedical research. Below are some typical application fields:

 

1. Disease Research

Itraq proteomics technology can be used to study the changes of proteins in different disease states. By comparing the abundance differences of proteins in healthy tissues and diseased tissues, potential biomarkers related to diseases can be discovered, providing a basis for early diagnosis and treatment of diseases.

 

2. Drug Development

Itraq proteomics technology can be used to evaluate the impact of drugs on the proteome. By comparing the abundance differences of proteins in the drug-treated group and the control group, the mechanism of action of the drug can be revealed, providing guidance for drug development.

 

3. Biological Research

Itraq proteomics technology can be used to study the changes of proteins in biological processes. By comparing the abundance differences of proteins at different time points or under different conditions, the molecular mechanisms of biological processes can be revealed.

 

Itraq proteomics technology reveals the molecular mechanisms of proteomics through its unique principles. It plays an important role in disease research, drug development, and biological research. With the continuous development of technology, itraq proteomics technology will provide more opportunities and challenges for us to understand proteomics more deeply.