Quantitative Proteomics
- High Sensitivity and Specificity: Capable of detecting and quantifying low-abundance proteins with high specificity and accuracy.
- High Throughput: Suitable for high-throughput analysis of large-scale samples, enabling rapid processing of numerous samples.
- Comprehensiveness: Allows simultaneous analysis and comparison of protein expression levels across multiple samples, providing comprehensive information.
Quantitative proteomics is a technique utilized for the quantitative analysis of protein expression levels to elucidate alterations in protein composition and function within cells, tissues, or organisms. This technique holds significant importance in biomedical research, aiding scientists in comprehending disease mechanisms, identifying biomarkers, and developing new therapeutic drugs. Quantitative proteomics fundamentally depends on mass spectrometry (MS) technology to accurately quantify proteins. The core principle involves analyzing protein samples with a mass spectrometer to obtain quantitative data by measuring the mass-to-charge ratio (m/z) of peptides and the intensity of fragment ions. Commonly employed methods include label-based and label-free approaches: (1) Label-based Methods: These methods involve the use of isotope labels or chemical tags to distinguish and compare different samples during mass spectrometry analysis. Techniques such as SILAC (Stable Isotope Labeling by Amino acids in Cell culture) and TMT (Tandem Mass Tagging) exemplify this approach. (2) Label-free Methods: These methods achieve quantification by directly comparing the intensity or area of peptide mass spectra across different samples. This approach simplifies experimental procedures and is suitable for large-scale sample analysis.
Quantitative proteomics is broadly applicable to the quantitative analysis of various proteins or peptides, especially those pivotal in disease research, drug development, and biomarker discovery. Examples include cell signaling proteins, metabolic enzymes, structural proteins, and immune-related proteins. MtoZ Biolabs offers the following quantitative proteomics services:
Label-Free Quantitative Proteomics
iTRAQ/TMT Quantitative Proteomics
SILAC/Dimethyl Quantitative Proteomics
SWATH Quantitative Proteomics
MRM Quantitative Proteomics
Chen, X. et al. Proteomics. 2015.
Figure 1. Characteristics and Applications of MS-based Quantitative Methods
Analysis Workflow
1. Sample Preparation: Proteins are extracted from cells, tissues, or organisms and subjected to enzymatic digestion (e.g., trypsin digestion) to generate peptides.
2. Labeling or Non-labeling Treatment: Depending on the chosen quantitative method, samples undergo isotope labeling or are directly analyzed by mass spectrometry.
3. Mass Spectrometry Analysis: Peptides are separated and detected using a mass spectrometer, generating mass spectrometry data.
4. Data Processing and Analysis: Specialized software is employed for the quantitative analysis and normalization of mass spectrometry data, comparing protein expression differences among samples.
5. Result Validation: The reliability of mass spectrometry results is verified using alternative methods such as Western blotting or ELISA.
Applications
1. Revealing Disease Mechanisms: By comparing protein expression levels between healthy and diseased states, the molecular mechanisms of diseases can be elucidated.
2. Discovering Biomarkers: Protein biomarkers indicative of disease diagnosis and prognosis can be identified.
3. Drug Development: The impact of drugs on protein expression is evaluated, optimizing drug design and efficacy assessment.
4. Biological Research: The functions and interactions of proteins within cells and organisms are revealed, advancing basic biological research.
Service Advantages
MtoZ Biolabs offers mass spectrometry-based quantitative proteomics services. Whatever your requirements may be, we are dedicated to meeting your needs. If you would like to inquire about our services, please feel free to contact us.
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