As a LifeSensors customer, your in vivo validation process, focusing on high-throughput assay development, begins with a consultation with members of our team. You will discuss target(s), project goals, assay format, and assay parameters. The consultation will determine how experiments such as cell culture, assay development, technology transfer, and drug treatment will be divided among the LifeSensors and you as a customer. After project completion, customer deliverables may include any or all of the following: assay development summary, sample proof-of-concept data, assay SOP, assay reagents, or drug activity data (e.g. cellular IC50s). Assay reagents may be off-the-shelf products or customized products to be sold exclusively to you as a customer. The project timelines for this process range from 4-8 weeks depending on assay format, target selection, and goals. LifeSensors will work with you to generate a timeline estimate for your individual project process.
By collaborating with LifeSensors for your drug discovery needs, you will get the data you need to validate your best compounds for advancement from in vitro hits to animal and clinical studies.
As a LifeSensors customer, we will schedule a consultation with you to discuss target(s), project goals, antibody selection, control samples and experimental conditions, for your UbiTest order. The consultation will determine whether cell culture and treatment may need to be performed either by a LifeSensors team member, or whether you will provide LifeSensors with treated cell samples to analyze. A LifeSensors scientist will then perform UbiTest assays here in our laboratory. Post-project deliverables will include western blot data indicating the ubiquitylation level in each of your samples. Project time estimation ranges are based on the number of samples to be analyzed .
1-2 weeks for 1-6 samples
2-3 weeks for 7-12 samples
For samples greater than 13, you must contact us for an estimated project time line
Your final UbiTest report will show the relative level of your protein’s ubiquitylation across each sample you submit for analysis. This will allow you to determine the downstream effect of inhibitors or biological stimuli on the ubiquitylation of your protein of interest. While ubiquitin Western blots are challenging, our data reports will allow you to draw convincing conclusions and accelerate your research.
High-Throughput Cellular Ubiquitylation Assays
Measuring cellular ubiquitylation in a plate-based format accelerates the process of ubiquitin pathway drug discovery. We have developed several platforms that enable rapid, quantitative monitoring of cellular ubiquitylation.
- LifeSensors’ UbiQuant™ S ELISA and AlphaLISA assays are industry-leading platforms for measuring ubiquitylation of a target protein in cells. These assays utilized TUBEs (Tandem Ubiquitin Binding Entities) to detect ubiquitylated substrates, and are individually optimized for a specific substrate protein.
LifeSensors continues to bring new technology to ubiquitin researchers, and we are proud to introduce the UbiQuant Ultra. This new generation of the UbiQuant family features Tandem Ubiquitin Binding Entities (TUBEs) coated plates for linkage-specific quantification. TUBEs bind with high affinity, ensuring ubiquitylated proteins are captured efficiently. UbiQuant Ultra plates can be coated with pan-selective, K48, K63, or M1 linear TUBEs depending or your research question. Please contact us to learn more about this exciting technology and receive custom pricing.
LifeSensors’ UbiQuant™ ELISA is intended for the accurate determination of the concentration of total ubiquitin (poly- + mono-) in cell and tissue lysates. This assay is useful for monitoring the effects of broad-spectrum compounds such as proteasome inhibitors on total ubiquitin levels.
UbiTest: Simple Assays for Cellular Ubiquitylation
The UbiTest platform replaces traditional immunoblotting methods for measuring cellular ubiquitylation, enabling simple comparisons of substrate ubiquitylation levels between samples and determination of ubiquitin chain linkage types. UbiTest is based on TUBE technology for enriching polyubiquitylated proteins. By visualizing the ubiquitylated fraction both with and without DUB digestion of polyubiquitin chains, UbiTest streamlines the confirmation of ubiquitylation status.
Previously, ubiquitin immunoblots have relied on the visualization of a qualitative, often deceptive smearing pattern, but no longer! Furthermore, the single band in the DUB-treated ubiquitlyated fraction simplifies comparisons between different samples, making this assay perfect for comparing the effect of biological stimuli or compound treatments on substrate ubiquitylation in several samples.
For more information about UbiTest, please also visit our product pages, where you can purchase off-the-shelf assay kits for performing UbiTest in your own lab.
LifeSensors’ High Throughput (HT) UbiTest is a customized medium-throughput assay for absolute quantification of ubiquitylated substrate levels in cells that combines UbiTest principles and TR-FRET technology.