VHL Protein Complex, Active, 10 µg

Quantitative assays for investigating ubiquitination pathways are essential for advancing ubiquitin-related drug discovery research. To illustrate the utility of in vitro ubiquitination enzyme cascades, we quantify the ubiquitination of Smac/DIABLO by the E3 ligase XIAP using a functional enzymatic reaction coupled to multiple assay readouts. We compare different detection methods that can be used to test small molecules for their potential to modulate XIAP-mediated ubiquitination in the context of this assay. The data provides a basis for use of these recombinant E3 ligase cascades in various applications, including screening campaigns.