SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture)-Based Quantitative Service
- Oncology: Analyzing protein expression heterogeneity in the tumor microenvironment, screening drug resistance biomarkers.
- Neurodegenerative Diseases: Tracking aggregation dynamics of misfolded proteins in brain tissue.
- Immunology: Quantifying activation levels of key proteins in inflammation signaling pathways.
- Drug Mechanism Studies: Evaluating the protein degradation effects induced by targeted drugs.
SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture) is an advanced protein quantification technique based on stable isotope labeling. By metabolically incorporating isotopically labeled amino acids (such as "light" and "heavy" isotopes) during cell culture, the entire protein profile of the cells can be labeled. The newly synthesized proteins in experimental and control groups can be distinctly differentiated in mass spectrometry based on mass differences, enabling precise quantification of protein expression, post-translational modification dynamics, and interaction networks.
Technical Principles
The core of SILAC lies in utilizing the cell’s dependence on isotope-labeled amino acids. Cells cultured in specific media fully replace natural amino acids with labeled amino acids, after at least five cell divisions, ensuring that all newly synthesized proteins incorporate these stable isotope-labeled amino acids. The experimental group ("heavy" labeled) and the control group ("light" labeled) are mixed, and the relative abundance of proteins is measured in mass spectrometry by comparing the "light/heavy" signal intensity ratio of the same peptide.
Chen, X. et al. Proteomics. 2015.
Figure 1. Workflow for Quantitative Proteomic Experiments Using SILAC
Technical Advantages
1. High Precision: Metabolic labeling avoids random errors introduced by sample handling.
2. Comprehensive Coverage: Compatible with complex samples like whole cell lysates and membrane protein-enriched samples.
3. Reproducibility: It significantly reduces batch effects and is suitable for high-throughput studies across multiple groups and time points.
SILAC is widely used in disease mechanism research, drug target screening, biomarker discovery, and other fields, especially in:
1. Low-abundance protein detection (e.g., phosphorylation signaling molecules).
2. Precise differential analysis (e.g., screening of proteins related to tumor resistance).
3. Multi-omics integration (combined transcriptomic/metabolomic analysis of regulatory networks).
Services at MtoZ Biolabs
With expert proteomics team and advanced mass spectrometry platform, MtoZ Biolabs offers end-to-end SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture)-Based Quantitative Service covering experimental design, cell culture, mass spectrometry analysis, and data interpretation, assisting in precise proteinomic research.
Our core service include:
1. Differential proteomic quantification: Detecting protein expression differences under various conditions (e.g., drug stimulation, gene editing, disease models).
2. Protein interaction network analysis: Using affinity purification techniques (AP-MS) to identify specific interacting proteins.
3. Post-translational modification (PTM) quantification: Quantifying modification sites like phosphorylation, ubiquitination, and acetylation.
4. Large-scale sample analysis: Supporting triple/five-way SILAC labeling for multi-group parallel experiments.
Analysis Workflow
1. Experimental Design & Labeling Strategy: Custom SILAC labeling plan design, including "light/heavy" label allocation.
2. Cell Culture & Label Verification: Culturing cells and confirming labeling efficiency via preliminary mass spectrometry.
3. Sample Preparation & Mass Spectrometry Analysis: Cell lysis → protein extraction and quantification → enzymatic digestion (Trypsin) → desalting purification; using nano-LC and high-resolution mass spectrometry for DDA/DIA data acquisition.
4. Data Analysis & Deep Mining: Protein quantification, differential protein screening, GO/KEGG enrichment analysis, protein interaction network visualization, modification site localization.
Why Choose MtoZ Biolabs?
1. High Sensitivity: Detect samples down to 10 ng, covering >5000 proteins/sample.
2. Flexible Customization: Compatible with mammalian cells, stem cells, primary cells, and some insect cell culture systems.
3. Fast Turnaround: Complete reports delivered in just 10-14 business days.
4. Comprehensive Data Interpretation: Provides raw data, visualized charts (volcano plots, heatmaps), and biological significance reports.
Applications
Case Study
1. SILAC-based proteomic quantification of chemoattractant-induced cytoskeleton dynamics on a second to minute timescale
This study utilizes SILAC-based proteomic methods to characterize rapid chemoattractant-induced dynamic changes in the actin–myosin cytoskeleton and regulatory elements on a proteome-wide scale, with second to minute timescale resolution. The approach provides insights into the kinetics of key cytoskeletal constituents and their association with both known and novel binding proteins. Validation is achieved through microscopy-based analysis of in vivo translocation dynamics for key signaling factors. This large-scale proteomic technique can be applied to other dynamic cellular processes. Our service offers high-resolution analysis of protein dynamics in response to various stimuli, providing insights into cellular processes such as cytoskeletal rearrangements and signal transduction. By using stable isotope labeling, we can accurately track protein abundance changes over time, allowing for detailed, time-sensitive investigation of cellular responses at a proteome-wide scale. This service is ideal for studies requiring high sensitivity and temporal resolution.
Sobczyk, GJ. et al. Nat Commun. 2014.
Figure 2. Heat-Map Representation of the Temporal Incorporation Profiles for the Cytoskeletal Proteins Detected in the SILAC Experiments
2. SILAC Mouse for Quantitative Proteomics Uncovers Kindlin-3 as an Essential Factor for Red Blood Cell Function
In this study, SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture) was used to label mice with either natural or 13C6-substituted lysine. Over four generations, the mice showed no significant changes in development, growth, or behavior. Mass spectrometry analysis revealed incorporation rates of proteins from blood cells and organs. In particular, the analysis of Kindlin-3-deficient erythrocytes showed structural defects in the red blood cell membrane skeleton. This approach provides a powerful tool for quantitative comparison of proteomes in knockout mice to study protein functions under in vivo conditions. Our quantitative proteomics services enable precise protein profiling through advanced labeling techniques such as SILAC. By utilizing mass spectrometry, we provide deep insights into protein expression and functional analysis, allowing for the comparison of knockout models and identification of key molecular players in various biological systems. This approach enhances the understanding of protein functions in complex in vivo environments.
Krüger, M. et al. Cell. 2008.
Figure 3. Analysis of b1 Integrin Knockout Platelets
With exceptional quantification accuracy and data reproducibility, SILAC technology is becoming a core tool for disease mechanism research, biomarker discovery, and drug development. MtoZ Biolabs, with its professional team, stringent quality control system, and efficient delivery capabilities, provides end-to-end SILAC quantitative solutions. Whether for basic research or translational medical needs, we collaborate throughout the process to ensure the maximum scientific value of your data. Contact us today for customized solutions and quotes!
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