Bradykinin B2R expressing cells labeled with Terbium

Tag-lite cells transiently expressing the Bradykinin B2 receptor labeled with Terbium for use in receptor binding applications. The cells are provided ready to use.
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  • Fully functional cell line
  • Transient expression
  • Ready for binding
Tag-lite cells transiently expressing the Bradykinin B2 receptor labeled with Terbium for use in receptor binding applications. The cells are provided ready to use.
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Overview

Bradykinin receptor has two known isoforms bradykinin type 1 and 2 receptors. Both receptors are GPCR but mediate different actions. While B1R signaling promotes local tissue inflammation by recruiting neutrophils, B2R signaling is believed to downregulate the proliferation and growth of cells in VSMC and cardiomyocytes.

Cells expressing the B2 receptor are provided pre-labeled with Terbium, and can be used to conduct receptor binding studies on the aforementioned receptor.

Assay principle

Running your B2 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 Bradykinin B2 labeled cells and their matching fluorescent ligand. Results may vary from one HTRF® compatible reader to another.
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