Our VHL binding assay kit is a fluorescence polarization (FP)-based assay that enables quantitative evaluation of ligand binding to the VHL E3 ubiquitin ligase complex (VBC). The VHL E3 ligase is part of the Cullin-RING ligase family, meaning it functions as a multi-protein complex. VHL acts as the substrate receptor, recognizing the target (physiologically HIF-1α) and binding to its adaptor proteins, ELOB/C, which connect it to the rest of the complex.

This kit uses CELT-504 as a high-affinity fluorescent tracer (low nanomolar range), allowing the determination of IC₅₀ and Ki values through competitive binding experiments in a robust binary format.

The assay has been validated using reference compounds such as VH298 and MZ1, providing affinity values consistent with reported data.

The homogeneous FP format allows direct monitoring of binding events without washing steps, making it suitable for compound screening, profiling and PROTAC development workflows.

For more information on fluorescence polarization techniques, please refer to our blog article.