Signal-Seeker® Ubiquitin Detection Kit (10 assays) - KIT 10 assays

Exceptionally sensitive and easy to use—capable of detecting endogenous ubiquitination even in low-abundance proteins such as c-Fos (1.35 × 10⁴ molecules per A431 cell).

Applications

• Detect endogenous ubiquitination in your protein of interest (POI)
• Discover novel regulatory mechanisms for your POI
• Confirm ubiquitination data generated through the transfected POI
• Confirm ubiquitination data generated through proteomic approaches
• Monitor PROTAC efficiency
• Discover novel biomarkers

Material

Kit contents (10 - 30 assays)

• BlastR® lysis buffer
• BlastR® dilution buffer
• BlastR-2® wash buffer
• De-ubiquitination/SUMOylation inhibitor
• Protease inhibitor cocktail
• Ubiquitination affinity beads
• Control beads
• Anti-ubiquitin-HRP antibody
• Precision Red™ advanced protein assay reagent
• Bead elution buffer
• DMSO
• Chemiluminescent detection reagents
• Spin column collection tubes
• BlastR® filters

Equipment & materials required

• Cell line or tissue
• Cell scrapers
• Liquid nitrogen for snap freezing lysates
• SDS-PAGE & Western blot equipment
• Primary antibody to POI
• Secondary antibody

Biological Activity Assay

The Signal-Seeker® ubiquitination detection kit uses affinity beads to enrich ubiquitinated (mon-, multi-, and poly) proteins from any given cell or tissue lysate. The enriched protein population is then analyzed by standard Western blot procedures, and the modified protein of interest is detected by the end-user using their primary antibody.

Key characteristics

• Sensitivity: Demonstrated to detect endogenous ubiquitination of multiple target POI
• Input requirements: 200 µg – 1 mg of lysate
• Timeframe: Approximately 10 hours, typically performed over two days