Ubiquitin is a small (8.6 kDa) regulatory protein and ubiquitination is an important post-translational modification, in which ubiquitins are attached to a substrate protein. This process contains three main steps: activation, conjugation, and ligation and requires three types of enzymes: ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s), and ubiquitin ligases (E3s).
Figure 1. The ubiquitin proteasome system (Suresh B, et al., 2016).
The ubiquitin-proteasome system mediates 80%-85% protein degradation in eukaryotes. In addition to participating in protein degradation, ubiquitination is involved in almost all life activities, including the cell cycle regulation, proliferation, apoptosis, differentiation, transcriptional regulation, gene expression, transcriptional regulation, signal transmission, damage repair, inflammatory immunity. Ubiquitination is related to the pathogenesis of diseases, such as cancer and cardiovascular diseases. Therefore, it has become a new target for research and development of new drugs.
Overview of Protein Ubiquitination Analysis
Creative Proteomics has already developed a professional platform for ubiquitin-proteomics analysis services to help you address research problems of ubiquitination identification and quantification. Through comprehensive protein ubiquitination analysis services, including protein extraction, digestion, peptide enrichment, LC-MS/MS analysis, data analysis and bioinformatics analysis, we can quickly, accurately and efficiently analyze the changes of various disease-related protein modifications to meet the needs of biomarker identification and drug target screening.
Figure 2. The workflow of protein ubiquitination analysis.
Sample Requirements
We can accept a variety of samples, including but not limited to:
- Plant tissues: roots, xylem, phloem, etc.
- Animal tissues
- Microorganisms
Body fluids: saliva, amniotic fluid, cerebrospinal fluid, serum, urine etc.
Protein extract. In order to ensure the test results, please inform the buffer components, whether it contains thiourea, SDS, or strong ion salts. In addition, the sample should not contain components such as nucleic acids, lipids, and polysaccharides, which will affect the separation effect.
If you want to know specific samples requirements, please feel free to contact us.
Delivery
- Detailed report, including experiment procedures, parameters, etc.
- Raw data and data analysis results (protein identifications and ubiquitin mapping)
Our Advantages
- High-throughput: identify and quantify up to thousands of proteins at once
- High precision and accuracy: accurate identification of modified peptides and modification sites
- 100% coverage: Use 2-3 enzymes for protein digestion to ensure full-coverage
- Ability to analyze low abundance ubiquitin-proteins
- State-of-the-art facilities: Triple TOF 5600, Q-Exactive, Orbitrap Fusion Tribrid
At Creative Proteomics, many excellent and experienced experts will optimize the experimental protocol according to your requirement and guarantee the high-quality results for protein ubiquitination. Creative Proteomics provides a broad range of technologies for ubiquitin research that enable quantification of protein amount and ubiquitination modification. Please feel free to contact to discuss your specific needs.
Reference
1. Suresh B, Lee J, Kim K S, et al. The importance of ubiquitination and deubiquitination in cellular reprogramming. Stem cells international, 2016, 2016.