Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful method to study the relative proteomic change under differential treatments, which relies on the mass spectrometry and the metabolic incorporation of amino acids with substituted stable isotopic nuclei. In SILAC, a given 'light' or 'heavy' form of the amino acid is incorporated into two samples. Two cell populations are grown in culture media that are identical except that one of them contains a 'light' and the other contains a 'heavy' form of a particular amino acid (e.g. 12C and 13C labeled L-lysine, respectively). As the two isotopically labeled amino acids are essentially chemically identical, their incorporation does not interfere with normal cell growth, while leading to proteins/peptides that are distinguishable by mass and thus are ideal for mass spectrometric analysis. SILAC approaches are well suited for monitoring changes in post-translational modifications.
Overview of SILAC-based Proteomics Analysis
At Creative Proteomics, we can provide a complete SILAC service, including cell culture, treatment of cells, and proteomics analysis. Based on SLIAC and mass spectrometry, we can analyze the relative proteomic change under differential treatments. In addition, compare with other technologies, we can provide protein-protein interaction and post-translational modification analysis. The ability to multiplex two or three samples per analysis allows for increased throughput and cost savings in quantitative proteomics experiments along with improved relative quantitation.
Workflow of SILAC-based Proteomics Analysis
- Cell culture and cell labeling.
- Sample preparation, including protein extraction, mixing unlabeled (light) and labeled (heavy) samples, trypsin digestion, peptide fractionation.
- LC-MS/MS analysis. An advanced platform equipped Triple TOF 5600, Q-Exactive, Orbitrap Fusion™ Tribrid™, etc.
- Data analysis by using professional software and database for reliable results.
- Detailed report.
Sample Requirement
We can accept not only non-labeled cell samples, but also samples at any stage after SILAC labelling.
- Non-labeled cell samples
- Labeled cell samples
- Protein samples
If you want to know specific samples requirements, please feel free to contact us.
Advantages of SILAC-based Proteomics Analysis Service
- High labeling rate: The labeling efficiency is not affected by lysate and efficiency can be as high as 100%.
- High sensitivity: The sample requirements are small, usually only tens of micrograms of protein per sample.
- High throughput: Mass spectrometry can identify and quantify hundreds to thousands of proteins simultaneously.
- High precision: Multiple samples are mixed, simultaneously digested, and identified. The subsequent experiments have the same effect on the sample, which reduces the impact of experimental operation and equipment, and has higher precision and reproducibility.
- High activity: The label uses in vivo labeling technology and is closer to the true state of the sample.
- Comprehensive services: Combined with other technologies, we can provide protein-protein interaction and post-translational modification analysis.
Bioinformatic Analysis:
To address customers’ problems, we provide bioinformatic analysis, including:
- Functional annotation and enrichment analysis
- Clustering analysis
- Network analysis
- Statistical analysis
Creative Proteomics provides an unbiased strategy that can reveal how specifically either inhibitors, or other perturbations affect the dynamic properties and cellular distributions of proteins. It can also be used as a sensitive and effective method to determine the specific interaction partners of proteins in the cell. Based on professional knowledge and experienced staff, Creative Proteomics provides all-inclusive SILAC services. As every project has different requirements, please contact our specialists to discuss your specific needs.
