Protein ubiquitination, a key regulatory process essential for life at the cellular level, triggers a wide range of functional consequences, such as targeting proteins for proteasomal degradation, facilitating protein-protein interactions, and altering subcellular transport events. In the last two decades, significant efforts have been made to identify ubiquitinated proteins through mass spectrometry (MS)-based proteomics studies. In these studies, modified peptide enrichment is an essential and critical step. Creative Proteomics focuses on post-translational modification (PTM) analysis. With years of experience in this area, our outstanding experts are able to provide effective and customized modified peptide enrichment services, including ubiquitinated peptide enrichment services. We have developed an efficient and robust workflow that can enrich ubiquitinated peptides directly from a pool of digested peptides, facilitating downstream MS-based identification of lysine ubiquitination sites from proteins.
PTM enrichment
The abundance of PTM is usually much lower and the occupancy of PTM sites is usually sub-stoichiometric. PTM enrichment processes are at the forefront of the field of proteomics, and the development of these methods allows researchers to continue to gather valuable information on dynamic changes that occur in the proteome. Phosphorylated peptides are usually enriched with immobilized metal affinity chromatography (IMAC) or titanium dioxide, acetylated peptides are mainly enriched with antibody K(Ac) against lysine acetylation, and ubiquitinated peptides are usually enriched with antibodies that recognize the Lys-ɛ-Gly-Gly (K-ɛ-GG), which is the residue of the ubiquitin side chain after trypsin digestion. In addition, peptide separation strategies such as strong cation exchange chromatography (SCX), hydrophilic interaction chromatography (HILIC) or reverse phase liquid chromatography (RPLC) are often employed to reduce the complexity and enhance the depth of coverage of PTM samples.
Ubiquitinated peptide enrichment service
We specialize in ubiquitination analysis of MS-based proteomics. To improve the sensitivity of protein ubiquitination identification, it is important to enrich ubiquitinated substrates from whole cell lysates to avoid interference from non-ubiquitinated peptides. Our ubiquitinated peptide enrichment service is primarily based on an anti-K-ε-GG antibody-based approach that specifically identifies lysine peptides containing ubiquitinated sites. Based on our robust workflow and experienced experts, we can greatly promote the sensitivity of ubiquitination analysis, enhancing the comprehensive mapping scale of ubiquitination signals. In addition to antibody-based enrichment strategies (the most widely used method for systematic analysis of ubiquitinated peptides), we are committed to developing other rapid, convenient and effective services to enrich for ubiquitinated peptides to meet the diverse needs of researchers.
Advantages of our service
- High specificity and affinity of ubiquitin antibodies.
- Effective enrichment and purification of ubiquitinated proteins.
- Detailed and comprehensive service procedures and results reporting.
- Dedicated after-sales service.
After digestion by various proteases, most peptides are non-ubiquitinated peptides, which significantly obscures the identification of ubiquitinated peptides. Based on our advanced platform and experienced experts, we are committed to providing customized and scalable ubiquitin peptide enrichment services to meet the diverse needs of our clients. If you are interested in our ubiquitinated peptide enrichment service, please feel free to contact us for details.
References
- Sun, Mingwei, and Xiaofei Zhang. "Current methodologies in protein ubiquitination characterization: from ubiquitinated protein to ubiquitin chain architecture." Cell & Bioscience 12.1 (2022): 1-17.
- Anania, Veronica G., et al. "Peptide level immunoaffinity enrichment enhances ubiquitination site identification on individual proteins." Molecular & Cellular Proteomics 13.1 (2014): 145-156.
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