Revvity, 64BDVHLPEG, HTRF Human VHL Binding Kit, 500 Assay Points

Overview
A fast and easy way to identify new binders to VHL protein.
VHL, also known as von Hippel-Lindau, is involved in many biological processes and is closely associated with the regulation of angiogenesis, cell growth, or cell survival.
VHL is one of the most popular E3 ligases recruited by bifunctional Proteolysis-targeting chimeras (PROTACs) to induce ubiquitination and subsequent proteasomal degradation of a targeted protein.
VHL interacts with several proteins to form the functional von Hippel-Lindau ubiquitination complex, in which VHL seems to act as a target recruitment subunit in the E3 ubiquitin ligase complex, and targets various proteins to proteolysis such as the hydroxylated hypoxia-inducible factor (HIF) under normoxic conditions.
Therefore, identifying new PROTAC VHL ligands can improve selective proteasomal-dependent degradation of proteins of interest, involved in the onset of diseases such as cancers.
How it works
Assay principle
The HTRF Human VHL Binding Kit is a competitive assay format which uses a VHL-Red Ligand as VHL ligand, a 6His-tagged human VHL protein complex, and an anti 6His Europium Cryptate-labeled antibody. VHL binding compounds compete with the VHL-Red Ligand and thereby prevent FRET from occurring.

 

Assay protocol
The VHL binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate. The 6His-tagged VHL protein complex is then added, followed by the dispensing of the HTRF reagents: The anti 6His antibody labeled with Europium cryptate and the VHL Ligand labeled with a Red HTRF acceptor. The reagents labeled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally. Application-Protein-Protein Interaction
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-5 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Binding Assay
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Inflammation Neuroscience Oncology & Inflammation
Unit Size-500 assay points