ELISA-Based In-Vitro Ubiquitylation Assay
Ubiquitylation is an important protein post-translational modification process in PROTAC® technology, and is the key to the degradation of the protein of interest (POI). BOC Sciences has established an advanced one-stop Proteolysis Targeting Chimeric (PROTAC) service platform to provide customers with ubiquitylation assay services, and enzyme-linked immunosorbent assay (ELISA) as a traditional technical means can quantify the ubiquitylation level of proteins. BOC Sciences has always been committed to providing high-quality research outsourcing services and has extensive experience in ELISA technology.
Introduction
Ubiquitylation refers to the process where ubiquitin specifically modifies the target protein under the action of a series of special enzymes. Ubiquitylation consists of three enzymatic reactions involving E1 ubiquitin activating enzymes, E2 ubiquitin conjugating enzymes, and E3 ubiquitin ligases. E1 ubiquitin activating enzymes activate ubiquitin and pass the activated ubiquitin to E2 ubiquitin conjugating enzymes. They promote the connection between their bound ubiquitin and POI through their active site cysteine. E3 ubiquitin ligases participate in ubiquitin transfer and promote the formation of β-peptide bonds between the target protein and the C-terminal carboxyl group of ubiquitin in the final stage. The protein substrate labeled with ubiquitin can be specifically recognized and degraded by the protease. Ubiquitinated proteins can be recognized by specific antibodies. ELISA is a qualitative and quantitative test of immune response using antibody-antigen specificity, and is a commonly used technical method in immunodiagnostic testing and biological science research. Anti-Ub antibody and ELISA technology can be used to quantitatively detect ubiquitinated protein, and then analyze the Ubiquitylation level of these proteins.
Our Services for ELISA-Based In-Vitro Ubiquitylation Assay (include but not limited to the following):
- Sandwich ELISA Assay
For sandwich ELISA to detect the concentration of ubiquitinated protein, the capture antibody is generally pre-coated on a 96-well plate, and then the samples are added for incubation. The ubiquitinated protein binds to the capture antibody, while the unbound analytes can be removed by washing process. The detection antibody can bind to the target protein and generate a specific detectable signal through some means. In addition to the traditional double antibody sandwich ELISA experiment service, BOC Sciences also provides other detection services based on double antibody sandwich ELISA. - Competitive ELISA Assay
Competitive ELISA measures the content of a certain analyte in the sample by quantifying the interference of the analyte. The signal intensity finally obtained is inversely related to the analyte content, so the higher the concentration of ubiquitinated protein in the sample, the weaker the signal obtained.
Our Advantages
- Experienced ELISA technical team.
- For different samples, the experimental operation of the differentiated testing scheme is carried out.
- Strict and standardized experimental operation process.
- Experimental results to ensure accuracy and repeatability.
- Professional data analysis.
- Complete test report, including real raw data, professional results analysis, and test related parameters such as equipment model, etc.
- Efficient testing services and competitive prices.
Workflow of ELISA-Based In-Vitro Ubiquitylation Assay
* PROTAC® is a registered trademark of Arvinas Operations, Inc., and is used under license.
