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Target Degradation

As an excellent service companion for your research discovery and development projects, Creative Biolabs offers one-stop PROTAC® service which can best fit your requirements and accelerate the progression of your project. The principle of PROTAC® is disease-causing target protein degradation. Herein, we provide different kinds of in vitro target degradation assays to detect quantitative degradation.

Background

The ubiquitin-proteasome system is a natural physical mechanism which is crucial in selective removal of target proteins. This system can finely tune the post-translation levels of target proteins which is very important to maintain the homeostasis. In UPS, ubiquitin can covalently link to a specific lysine of the target protein. The consequence of the polyubiquitination is high molecular weight protein-ubiquitin complex which can be recognized for degradation by 26S proteasome. This process is mediated by a series of enzymes including E1, E2, and E3. PROTAC®s contains specific E3 ligase ligand through which the target proteins can be ubiquitylated and ultimately degraded. Hence, verifying the target protein degradation is essential to evaluate the function of designed PROTAC®s.

Introduction of Target Degradation Assays

The degradation of disease-causing proteins is the final goal of PROTAC®s. Target ubiquitination plays an important role to indicate the potential degradation efficacy and is a direct way to evaluate the degradation efficacy. Creative Biolabs offers 3 different categories of target degradation assays for your flexible choice, including western blot, reporter gene assay, and custom downstream effects.

  • Western blot
  • WB is a typical classic technique in cell and molecular biology. By using the specific anti-target protein antibodies, we can identify the post-transcription protein expression level easily from a complex mixture of proteins extracted from the cells. We offer proper primary and secondary antibodies. As a leading antibody supplier, Creative Biolabs owns a fantastic antibody library and also custom antibody production service in case the catalog antibody can’t fit your projects. In addition, we provide target protein plasmid and stable cell line construction service. Foremost, we established a mature WB platform from cell culture, assay design, protein extraction, gel electrophoresis, staining, to immunofluorescence, etc.

    Schematic diagram of western blot. Fig.1 Schematic diagram of western blot.

  • Reporter gene assay
  • Reporter genes can be utilized to study the gene expression level easily at a real-time level. They are constructed into the cells, and the expression level of the reporter gene can reflect the expression level of target proteins via a direct assay or monitoring the reporter protein enzymatic activity. Creative Biolabs provides 3 reporter gene assays for your flexible choice, including green fluorescent protein (GFP), luciferase, and blue-white screen.

    Schematic diagram of reporter gene assay. Fig.2 Schematic diagram of reporter gene assay.

  • Downstream effects
  • Except for the direct way to detect the target protein degradation, downstream effects of target protein can also be detected to evaluate the PROTAC® efficacy. Creative Biolabs can design the custom assay for you to detect the downstream effects based on the requirements of your projects. With high-level scientists, we ensure the optimal downstream signal targets are designed to fit your projects.

Highlight Features of Our Services

  • Accurate quantitive analysis for both WB and reporter genes
  • Short turnaround time and high-quality data
  • Both direct and indirect assays available for your flexible choice
  • Extensive collaboration experience with numerous worldwide clients

With high-level scientists and skillful technicians, Creative Biolabs has blossomed into a service leader in drug discovery and development field. We are committed to providing cutting-edge services to our clients at the forefront of technological innovation. Please contact us for more detailed information. We are always here to provide you with the best technique.

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