Revvity, L0025RED, Tag-lite Opioid Receptor (Orexin-A) Red Agonist Ligand

Tag-lite receptor binding assays use either a green or a red fluorescent ligand to function. This Orexin receptor red agonist is an OrexinA derivative labeled with a red emitting HTRF fluorescent probe. This ligand is best suited for use in the Orexin 1 and Orexin 2 receptor binding assays. Tag-lite assays offer many features and applications over traditional receptor/ligand binding assays: Non-radioactive Homogeneous and filtration-free Ready-to-use kits and reagents for binding assays Does not alter the receptor pharmacology Peer-reviewed, validated technology Thorough kinetic studies with true Kd and Ki values, association (Kon) and dissociation (Koff) rate constants Screening and profiling of biologics and large molecules
Assay principle Running a receptor binding assay using Tag-lite is as easy as it can get. Simply dispense 10 µL of labeled cells into each well, followed by 5 µL of labeled ligand and 5 µL of the compound you wish to test. Like all HTRF assays, Tag-lite assays do not require any washing steps. A diagram of the procedure to be followed is given on the right. Saturation binding (KD) A saturation binding assay measures total and non-specific binding for increasing concentrations of ligand under equilibrium conditions. To perform the assay, the fluorescent ligand is titrated into a solution containing a fixed amount of labeled cells and then incubated to equilibrium. The HTRF ratio obtained from this titration is the total binding. Competitive binding (KI) A competitive binding assay is performed to measure the dissociation constant, Ki. To perform the assay, the compound is titrated into a solution containing a fixed concentration of fluorescent ligand and a fixed amount of cells.