SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture) is a breakthrough technology that has significantly influenced the field of quantitative proteomics. As a popular method for its high accuracy and reproducibility, this technique has been widely used for accurate fractionation to measure differential changes in the relative abundance of proteins within a single protein sample. Our team at BOC Sciences takes advantage of more than 20 years of experience to offers in depth SILAC services, empowering research in different biological domains.
What is SILAC Analysis?
SILAC involves metabolically labeling the proteome of cells in culture by incorporating stable isotope labeled amino acids. For the quantitative comparison of protein expression levels between conditions this is a necessary step. Every time a new protein is synthesized, SILAC incorporates isotopically-labeled amino acids in lieu of natural amino acids, so the development of each new protein possesses those labels, for accuracy in mass spectrometry quantitation techniques.
How Does SILAC Labeling Work?
SILAC is a metabolic labelling technique in which cells or organisms are grown in a medium containing light or heavy amino acids; incorporation of these amino acid into the proteome of the cells enables the distinction of proteins based on level of synthesis in two cell populations. At the core of SILAC stands the labeling of amino acids with stable isotopes. These isotopes, which includes carbon-13 (13C) or nitrogen-15 (15N) are non-radioactive and do not change the chemistry of the amino acids. These are also amino acids with a high propensity to be incorporated in to tryptic peptides so they can be easily detected in the MS. Even it has many merits like high preciseness, thoroughly physiological relevance, comprehensive proteome-wide coverage etc. which makes it a big gold standard in proteomics research.
What is the Difference Between SILAC and pSILAC?
SILAC directly labels proteins in the proteome as they are being synthesized by the cell, due to incorporation of isotopically labelled amino acids into growing proteins, while pSILAC provides information about protein degradation and synthesis rates. Using a short pulse of labeled amino acids, pSILAC allows researchers to better quantify rates of dynamic processes like protein degradation and synthesis.
| SILAC | pSILAC |
|---|
| Overview | A Continuous Amino Acid Labeling Approach | A Pulsed Amino Acid Labeling Approach |
| Mechanism | In SILAC, cells are cultured in media containing stable isotopically labeled amino acids for an extended period. The labeled amino acids are incorporated into newly synthesized proteins during cell growth and division. | pSILAC involves a short pulse of isotopically labeled amino acids followed by a chase period with unlabeled amino acids. This pulse-chase labeling strategy allows researchers to track newly synthesized proteins over time. |
| Labeling Strategy | Continuous labeling approach with stable isotopes. | Pulsed labeling approach with a short labeling period followed by a chase period. |
| Experimental Design | Suitable for studying steady-state protein expression levels and long-term changes. | Ideal for investigating dynamic protein synthesis rates and turnover kinetics. |
| Applications | Mainly used for quantifying relative changes in protein abundance under different conditions. | Particularly useful for studying dynamic processes such as protein turnover and synthesis rates. |
Professional SILAC Analysis Platform
Our SILAC services are powered by an advanced technology platform at BOC Sciences.
- High-Resolution Mass Spectrometers: Instruments such as the Triple TOF 5600 and Q-Exactive provide unparalleled accuracy and sensitivity.
- Specialized Media: We utilize ready-to-order media designed to lack a particular amino acid, and then provide back the > 90% isotopically labeled form to make complete incorporation.
- Dialyzed Serum: We use dialyzed serum in our culture media to avoid contamination by unlabeled amino acids.
Diversified SILAC Products
Diversified SILAC Services
Features of Our SILAC Analysis Platform
- In Vivo Labeling: SILAC labels proteins in their natural cellular context, preserving physiological relevance.
- High Labeling Efficiency: Our protocol achieves over 99% labeling efficiency.
- Minimal Experimental Variability: Incorporating labels during cell culture reduces experimental errors.
- High Sensitivity and Precision: Requires minimal protein amounts and provides a wide linear range for accurate quantification.
- Comprehensive Coverage: Capable of identifying and quantifying thousands of proteins simultaneously.
SILAC Analysis Service Content
Culture Medium Preparation
Customized culture media that we prepare for your needs, optimal growth conditions and totally labeled.
Isotope Labeling and Efficiency Detection
Our experts handle the labeling process and verify the incorporation efficiency using advanced mass spectrometry techniques.
Protein Extraction and Analysis
We extract proteins from labeled cells, perform SDS-PAGE separation, and analyze the samples via LC-MS/MS. Our service includes protein identification, quantitative analysis, and differential expression screening.
Bioinformatics Analysis
Our bioinformatics team provides comprehensive data analysis, including protein interaction networks, pathway analysis (e.g., GO, KEGG), and visualization tools such as volcano plots and PCA.
Detailed Reporting
Clients receive a complete report detailing experimental procedures, parameters, raw data, and analysis results.
SILAC Analysis Service Advantages
Expertise and Experience
With over 20 years in the industry, BOC Sciences brings unparalleled expertise and experience to every project, ensuring reliable and high-quality results.
Tailored Experimental Design
We offer customized SILAC protocols to meet your specific research needs, optimizing experimental routes for the best outcomes.
Competitive Pricing
Our services are competitively priced, providing high-value solutions without compromising on quality.
Comprehensive Support
From technical consultation to after-sales support, we are committed to assisting our clients throughout their research journey.
Applications of SILAC
Drug Target Research
SILAC is invaluable in identifying potential drug targets by analyzing changes in protein expression and interactions under different treatment conditions.
Dynamic Protein Studies
Researchers use SILAC to study dynamic changes in protein expression, synthesis, and degradation in various biological contexts, including cell differentiation and stress responses.
Disease Marker Screening
SILAC facilitates the identification of biomarkers for diseases by comparing protein expression profiles in healthy and diseased states.
Molecular Interaction Analysis
Quantitative analysis of protein-protein, RNA-protein, and DNA-protein interactions helps elucidate complex biological networks and pathways.
Post-Translational Modifications
SILAC enables the quantitative screening of post-translational modifications such as phosphorylation, methylation, and acetylation, crucial for understanding regulatory mechanisms in cells.
Customer Notice
Technical consultation
Before starting the experiment, we will wholeheartedly provide you with an effective SILAC experimental design plan. We can provide stable isotope-labeled amino acids.
Sample requirements
The customer provides the sample and sample information to be identified, and the sample should avoid all kinds of pollution and repeated freezing and thawing.
BOC Sciences' SILAC Analysis Services provide a robust and precise method for quantitative proteomics, supporting a wide range of applications from drug discovery to biomarker identification. Our professional team and advanced technology platform ensure high-quality, reliable results tailored to meet the specific needs of your research. Whether you are studying protein dynamics, interactions, or modifications, our SILAC services are designed to deliver comprehensive and accurate insights, driving forward the frontiers of proteomic research.
FAQ
1: What is the difference between SILAC and TMT?
Protein Quantification with SILAC (Stable Isotope Labeling by Amino acids in Cell culture) and TMT (Tandem Mass Tags) SILAC involves growing cells in culture in medium containing heavy isotopically labeled amino acids for several cell divisions, thus generating populations of labeled proteins that can be compared to unlabeled proteins in other related biological samples. Alternatively, TMT utilizes chemical tags to mark peptides generated from different samples which permits proteome wide quantitation across multiple conditions or time points in a single experiment. Finally, TMT is capable of quantifying proteins derived from complex samples such as tissues or body fluids.
2: Why is SILAC used?
SILAC Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) is a label for acurate quantification of protein expression and protein turnover in proteome scale. It is useful to compare protein levels in separate conditions or treatments in cell culture experiments. SILAC enables accurate quantitation of proteins and reduces measurement variability, facilitating the detection of minor changes in protein levels.
3: Is SILAC radioactive?
No, SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture)SILAC is not radioactive. Stable isotopes of amino acids (13C, 15N,or 2H) dully modify proteins, unlike in the radioactive pattern. This labelling approach permits the precise quantitation of proteins with no need to add radioactivity to the experiment.
4: What is TMT in proteomics?
TMT (Tandem Mass Tag) is a chemical labeling that is used to quantify and compare the levels of protein expression in several samples in proteomics. It consists of labeling peptides from multiple samples with isobaric mass chemical labels. Once labeled, the samples may be pooled for screening together in a single mass spectrometry experiment. TMT allows for the accurate and high-throughput quantification of proteins, which is key to understanding biological processes and disease.
5: What are the methods of protein Labelling?
There are basically three types of tags in use: stable isotopes, mass tags and fluorophores. All three tag types are widely used for protein labeling in diverse settings. Quantitative proteomics uses stable isotopes, Isotope-labeled forms of amino acids or other molecules that are added to proteins. Multiplexed protein quantification by mass spectrometry using mass-tags TMT and iTRAQ. In contrast, fluorophores allow proteins to be visualized (fluorescence microscopy, flow cytometry) or detected. These each have benefits to researchers based on the type of tag.
