|Description:||The TR-FRET Vitamin D Receptor (VDR) Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential VDR ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a VDR ligand-binding domain (VDR-LBD) that is tagged with glutathione-S-transferase (GST), in a homogenous mix-and-read assay format.
To run the TR-FRET VDR Coactivator Assay, VDR-LBD is added to ligand test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti-GST antibody. After room temperature incubation, the TR-FRET 520: 495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based upon the biology of the VDRcoactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide.
|Applications:||Co-Factor Interaction Assay|
|Storage:||Store in ultra-cold freezer (-68 to -85°C).|
|Size:||800 x 20 µL assays|
|Kit Components:||The TR-FRET VDR Coactivator Assay Kit contains the VDR-LBD (GST) protein, fluorescein-labeled TRAP220/DRIP-2 coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).|
|Materials Required but Not Supplied:||Microplate Reader|