The Ubiquitin Proteasome System (UPS) is a dynamic process involved in various cellular functions, including cell signaling, DNA repair, and proteasomal degradation. Ubiquitin can form diverse chains on its seven lysine residues, with K48 and K63-linked polyubiquitination being a crucial post-translational modification. While K48 polyubiquitination predominantly drives protein degradation and maintains cellular homeostasis, K63 linked polyubiquitination plays a pivotal role in various cellular processes, primarily associated with trafficking, signaling pathways, DNA repair mechanisms, and the regulation of protein-protein interactions. LifeSensors’ K63 Ubiquitin Linkage ELISA Kit enables relative and absolute quantitation of K63 ubiquitination on target proteins in cellular and tissue lysates under various treatments and conditions.
New assay plate developed for colorimetric detection allows for quantitative measurement using absorbance-based plate readers. Colorimetric readouts (650nm, 480nm) are more consistent day-to-day than luminescent-based detection methods, fast, and easy to use.
This assay replaces labor-intensive, semi-quantitative Western blot methods for examining K63 polyubiquitination, offering quantitative, reproducible results and facilitating high-throughput screening for compound library processing and structure-activity relationship (SAR) studies in drug discovery applications.
*The plate is designed for research use only.
PROTAC Assay Kit Components:
| Coated plate: |
One pre-coated and blocked 96-well strip plate (Store at -80°C) |
| Blocking concentrate (BC): |
12mL of 5X blocking agent for antibody dilution (Store at 4°C) |
| Plastic plate seals: |
Two plate seals provided |
| Colorimetric reagent (CR): |
5.0mL vial of CR reagent (Store at 4°C) |
| Stopping reagent (SR): |
5.0mL vial of SR reagent (Store at 4°C) |
| UPS Inhibitor Cocktail (10x) |
1000µL vial (in RIPA Lysis Buffer; Store at -20°C)(Contains PR-619, MG-132 and 1,10-phenanthroline) |
| Proteasome Inhibitor (10x) |
1000µL vial (in RIPA Lysis Buffer; Store at -20°C) |
