Post-translational modifications (PTMs) controlling various biological functions are a key aspect of protein diversity. To help clients achieve reliable and comprehensive PTM analysis, Creative Proteomics offers different mass spectrometry (MS)-based strategies, including bottom-up, middle-down, and top-down proteomics. Among them, the bottom-up approach focusing on the peptide level is the commonly used method with high sensitivity and high-throughput. By optimizing the protein extraction processes, peptide enrichment strategies, MS acquisition methods, and data analysis, our team of experts is dedicated to providing the most robust and accurate bottom-up MS-based PTM analysis services.
Bottom-up MS-based proteomics
MS-based methods are currently the gold standard and most informative technique for the analysis of proteins and their PTM. To date, a variety of MS-based techniques have been applied, including bottom-up, middle-down, and top-down approaches. The bottom-up approach is characterized by the digestion of proteins into short peptides (usually 500-3000 mass range) prior to analysis. Usually, after digestion with serine protease or trypsin, a specific tag can be introduced into the molecule so that the specific modification reacts with the chemical tag, which can then be analyzed with a reporter ion. It is worth pointing out that bottom-up MS-based proteomics is suitable for peptide specific modification analysis and determination of peptide modification status, but has limitations in PTM mapping and identification of alternatively spliced protein variants.
Advantages of top-down proteomics
- Well-developed for protein quantitation.
- Robust, sensitive, and high-throughput for analyzing small peptides and their PTMs.
- Dependent on bioinformatics tools for protein sequence determination.
- Suitable for peptide-specific modification analysis and determination of peptide modification status.
Bottom-up MS-based post-translational modification analysis service
Based on advanced instruments and experienced experts, we have established a robust and effective MS-based bottom-up proteomics workflow for protein PTM analysis. Since protein PTMs are characterized by substoichiometric, transient, and labile, we combine additional experimental steps to improve proteomics techniques for PTM analysis. For example, enrichment of the PTM of interest is often performed prior to MS analysis. To accelerate the success of our customers' projects related to multiplex PTM analysis, we offer a range of enrichment strategies to reduce sample complexity and improve the efficiency and reliability of the analysis, such as strong cation exchange chromatography (SCX), hydrophilic interaction chromatography (HILIC), immobilized metal affinity chromatography (IMAC), and titanium dioxide (TiO2)-based enrichment. In particular, we have expertise in phosphopeptide enrichment, acetylated peptide enrichment, ubiquitinated peptide enrichment, glycopeptide enrichment, and histone isolation and enrichment, and provide related services. In addition, our service combines multimodal LC-MS/MS and developed bioinformatics tools for various PTM analysis.
Major benefits of our service
- Advanced proteomics platform to ensure deep coverage.
- Experienced professionals work with the highest standards.
- Providing robust and reproducible results for future follow-up studies.
- Detailed analysis process and easy-to-follow reports.
Related services
Feel free to contact us for more information about our MS-based PTM analysis services. We look forward to working with you on your next project.
Reference
- Virág, Dávid, et al. "Current trends in the analysis of post-translational modifications." Chromatographia 83.1 (2020): 1-10.
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