Revvity, 64BDMDM2PEH, HTRF MDM2 Binding Kit, 10,000 Assay Points

Overview
A fast and easy way to identify new binders to MDM2 protein.
MDM2, also known as murine double minute 2, is involved in many biological processes and is closely associated with DNA repair, cell cycle arrest, apoptosis, and the occurrence of many diseases. MDM2 is one of the most popular E3 ligases which is recruited by bifunctional Proteolysis-targeting chimeras (PROTACs) to induce ubiquitination and subsequent proteasomal degradation of a targeted protein. MDM2 interacts with several proteins to form the functional E3 ubiquitin ligase complex, in which MDM2 functions as a substrate receptor of the E3 ubiquitin ligase complex and targets various proteins to proteolysis.
Therefore, identifying new PROTAC MDM2 ligands can improve selective proteasomal-dependent degradation of proteins of interest, involved in disease onset such as cancers, neurodegenerative, and metabolic disorders.
How it works
Assay principle
The HTRF MDM2 Binding Kit is a competitive assay format which uses MI-1061 Red Ligand as MDM2 ligand, a GST tagged human MDM2 protein, and an anti GST Europium Cryptate-labeled antibody. MDM binding compounds compete with the HTRF MI-1061 Red Ligand and thereby prevent FRET from occurring.

 

Assay protocol
The MDM2 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 plateT the MDM2 GST-tagged protein is then added, followed by the dispensing of the HTRF reagents: the anti GST antibody labeled with Europium cryptate and the MI-1061 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
Technology-TR-FRET
Therapeutic Area-Inflammation Neuroscience Oncology & Inflammation
Unit Size-10,000 assay points