Relative Protein Quantitative Service, MS Based

Relative quantification of proteins is a MS-based technique that compares the relative proteins expression levels between different samples, analyzing differential protein expression under different conditions and revealing the role of proteins in biological processes. Commonly used methods include stable isotope labeling by amino acids in cell culture (SILAC), isobaric tags for relative and absolute quantification (iTRAQ), and tandem mass tags (TMT). They achieve relative quantification by enzymatic digestion, isotopic labeling, separation, and mass spectrometric analysis.

MtoZ Biolabs offers protein relative quantification services based on Thermo Fisher's Orbitrap Exploris 240 mass spectrometry platform combined with nanoLC-MS/MS nanoscale chromatography, using multiple relative quantification techniques such as isotopic labeling (e.g., SILAC, TMT, iTRAQ) and label-free methods (e.g., SWATH-MS, DDA). We precisely compare protein expression levels across different experimental conditions or biological samples, identify protein-level changes, and help clients rapidly and accurately assess differences in protein expression levels to reveal the molecular mechanisms of biological processes, disease onset, and drug therapy.

Analysis Workflow

1. Sample Preparation

Proteins extraction from different biological samples using suitable methods according to project requirements, followed by purification, concentration, and quality assessment to ensure the sample meets experimental requirements.

2. Protein Digestion

Protein digestion by various enzymes into peptide for subsequent mass spectrometry analysis.

3. Isotopic Labeling

Peptide labeling by isotopic labeling methods such as SILAC, iTRAQ, or TMT.

4. Peptide Separation

Peptide separation by HPLC (e.g., RP-HPLC).

5. Mass Spectrometric Analysis

Analysis and detection of peptides by high-resolution mass spectrometry to obtain data on mass, m/z and other information.

6. Data Analysis

Analysis of mass spectrometry data by specific bioinformatics software to identify and relatively quantify proteins to find significantly different proteins by comparing differential protein expression under different conditions.

Service Advantages

1. High-Throughput Data Acquisition

Advanced Orbitrap Exploris 240 mass spectrometer with high-sensitivity, high-resolution protein detection to quickly acquire large amounts of protein data.

2. Accurate Quantitative Analysis

Efficient data processing workflows and software for accurate and reliable protein quantification.

3. High Sensitivity

Ability to detect low-abundance proteins with high sensitivity.

4. Broad Applicability

Suit for a wide range of biological samples, including cells, tissues, and body fluids to study various biological processes.

5. Customized Analytical Solutions

One-stop services including experimental design, sample processing and data analysis according to project requirements.

Applications

1. Gene Expression Regulation Studies

Compare protein expression differences under different conditions to study the effects of post-transcriptional regulation and post-translational modifications on gene expression.

2. Signal Transduction Pathway Studies

Use quantitative analysis of differential protein expression of signal molecules to reveal activation and inhibition mechanisms of signaling pathways.

3. Comparative Proteomics Studies

Compare protein expression differences among species or subtypes to explore genetic variation, evolutionary relationships, and functional differences in species.

4. Protein Interaction Network Studies

Reveal protein interactions and construct protein interaction networks by quantifying protein expression differences and correlations.

5. Disease Diagnosis and Molecular Mechanism Research

Identify potential disease-related biomarkers and delve into the molecular mechanisms of disease onset by comparing protein expression differences between normal and pathological samples.