Angiotensin II receptor type 2 or simply AT2 is a GPCR expressed during fetal development. AT2 is involved in vasodilation, inhibition of cell growth, and possibly natriuresis.
Cells expressing the AT2 receptor are provided pre-labeled with Terbium, and can be used to conduct receptor binding studies on the aforementioned receptor.
PRE-LABELED WITH TERBIUM
FROZEN CELLS, READY TO BE USED
CELLS/LIGAND VALIDATED FOR BINDING
Running your AT2 receptor binding assay using Tag-lite is as easy as it can gets. 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.
Watch this video explaining how to run a saturation binding assay using Tag-lite.
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.
Watch this video explaining how to run a competitive binding assay using Tag-lite.
Kd and Ki validation
Examples of data obtained using Angiotensin AT2 labeled cells and their matching fluorescent ligand (L0007RED). AT2 was used as reference ligand. Results may vary from one HTRF® compatible reader to another.