• Detect endogenous ubiquitination changes on a protein of interest (POI) in cells or tissue extracts
• Isolate the total ubiquitinated protein profile from cells or tissues via immunoprecipitation (IP) for downstream proteomic analysis
• Confirm POI ubiquitination data generated from mass spec proteomics

Protein ubiquitination is a post-translational modification where ubiquitin molecules are covalently attached to target proteins, regulating their degradation, location, or activity, with dysregulation linked to diseases such as cancer, neurodegeneration, and immune disorders. This pathway is also harnessed in PROTAC (Proteolysis Targeting Chimera) technology, which selectively induces the degradation of disease-related proteins for therapeutic benefit.

UBA01B-beads are an affinity reagent comprising a high-affinity/specificity TUBE (Tandem Ubiquitin-Binding Entities) protein covalently linked to agarose beads.

Key characteristics

• Proprietary TUBE formulation captures mono-, multi-, and poly-ubiquitinated proteins
• Broad spectrum enrichment of polyubiquitin species, including K48, K63, and linear chains
• Sufficient reagent for 40 IP assays (1 mg lysate per IP)
• Consistent lot-to-lot performance
• Supplied as a lyophilized powder
• Buffer composition after rehydration: 80 mM PIPES pH 7.2, 4% sucrose, and 4% polyethylene glycol

The biological activity of UBA01B-beads is demonstrated by their ability to enrich a broad range of ubiquitinated proteins from HeLa cell lysates (20 µl beads per 1 mg lysate). Specificity is shown by detecting ubiquitinated Rac1 following cytotoxic necrotizing factor 1 (CNF1) treatment, which is known to induce Rac1 ubiquitination. In this system, the Ub–Rac1 signal increased by at least 50-fold compared to untreated lysates (see datasheet).