Label-Free quantitative proteomics technology involves mass spectrometry analysis of protein-digested peptides using liquid chromatography-mass spectrometry (LC-MS) without the need for expensive stable isotope labels as internal standards. It simply analyzes the mass spectrometry data generated during large-scale protein identification to compare the signal intensities of corresponding peptides in different samples, thus achieving relative quantification of the proteins corresponding to the peptides.
Based onSILACandiTRAQQuantitative proteomics research technologies like SILAC and iTRAQ first label proteins and then analyze them using mass spectrometry. Although they provide extensive protein quantification information, the labeling reagents are relatively expensive, making each sample costly. Therefore, these methods are more suitable for quantitative analysis of protein expression changes at the whole proteome level. For relatively simple protein mixtures, such as co-IP samples, the cost-effectiveness is low. In the case of certain post-translational modification studies, such as ubiquitination, which occurs on lysine side chains, the enrichment of ubiquitinated peptides is achieved using antibodies against the two glycine (gly-gly) residues remaining on the lysine side chain. However, the iTRAQ labeling can significantly affect this enrichment process, making it unsuitable for analyzing ubiquitination modifications at the proteomic level using iTRAQ labeling techniques. Label-Free quantitative analysis technology was developed to meet these experimental needs. Since the ionization efficiency of the same peptide is consistent during mass spectrometry analysis, the area of the mass spectrometry peak directly represents the amount of the peptide, allowing direct determination of the protein's relative quantification information by comparing the mass spectrometry peak areas of the same peptide.
Biotec Pac employs Thermo Fisher's Q Exactive HF, Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometry platforms combined with Nano-LC to offer a Label-Free quantitative proteomics analysis service package. You only need to tell us your experimental objectives and send us your cells; we will take care of all subsequent project matters, including cell culture, cell labeling, protein extraction, protein digestion, peptide separation, mass spectrometry analysis, raw data analysis, and bioinformatics analysis.
Label-Free Quantitative Proteomics Service Process
Regarding Samples
1. If you provide co-IP samples, the required protein amount is around 2-5ug. Please do not include detergents (e.g., SDS) or high concentrations of oxygen in the sent protein samples. Alternatively, you can send us the beads containing the protein samples, and we will handle the subsequent experiments.
2. If you provide tissue samples, please send them to us on dry ice. Animal samples should be over 200mg, blood samples at least 1ml (ensure plasma is anticoagulated with EDTA), serum 0.2-0.5ml, urine 2ml, plant samples at least 1g, cell samples should be 1X10^7 cells, yeast or microbial samples should have a dry weight of 200mg.
3. If you provide protein samples, please ensure a total protein amount of over 50ug; ordinary tissue or cell lysis buffer can be used for protein extraction.
4. Sample transportation: Please use sufficient dry ice for transport and choose a faster postal method to minimize the possibility of sample degradation during transportation.
Bilingual Project Report
In the technical report, Biotec Pac will provide you with a detailed bilingual version (Chinese and English) technical report, including:
1. Experimental Procedures (Chinese and English)
2. Relevant Mass Spectrometry Parameters (Chinese and English)
3. Mass Spectrometry Images
4. Raw Data
5. Protein Differential Level Analysis
6. Bioinformatics Analysis
Label-Free Quantitative Proteomics One-Stop Service
You only need to place an order and send samples
Biotec Pac's one-stop service completes: Sample processing - Instrument Analysis - Data Analysis - Project Report
