Adenosine A3 receptor expressing cells labeled with Terbium

Tag-lite cells transiently expressing the A3 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 Fully functional cell line
  • Transient expression Transient expression
  • Ready for binding Ready for binding

Tag-lite cells transiently expressing the A3 receptor labeled with Terbium for use in receptor binding applications. The cells are provided ready to use.

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Overview

A3 receptor is one of the four adenosine receptor family (including A1, A2A, and A2B receptors). A3 receptor is implicated in various pathological conditions, such as cardiac and cerebral ischaemia, and neurodegenerative diseases, as well as inflammatory pathologies including rheumatoid arthritis and asthma. Thus, it is considered as an interesting therapeutic target.

Cells expressing the A3 receptor are provided pre-labeled with Terbium, and can be used to conduct receptor binding studies on the aforementioned receptor.

Benefits

  • PRE-LABELED WITH TERBIUM
  • FROZEN CELLS, READY TO BE USED
  • CELLS/LIGAND VALIDATED FOR BINDING

Assay principle

Running your A3 receptor binding assay using Tag-lite is as easy as it can get. Simply dispense 10 µL of labeled cells (C1TT1A3) into each well, followed by 5 µL of labeled ligand (L0069GRE) 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.

Diagram of Adenosine A3 receptor binding assay using the Tag-lite protocol

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.

Diagram of a saturation binding assay using Tag-lite
Representative data obtained when running a saturation binding assay

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.

Diagram of a competitive binding assay using Tag-lite
Representative data obtained when running a competitive binding assay
Watch this video explaining how to run a competitive binding assay using Tag-lite.

Kd and Ki validation

Examples of data obtained using Adenosine A3 receptor labeled cells and their matching fluorescent ligand (L0069GRE). XAC (Xanthine amine congener) was used for Ki determination in the competitive binding assay. Results may vary from one HTRF® compatible reader to another.

kd determination using A2B labeled cells and their matching fluorescent ligand L0068RED
ki determination using A3 labeled cells and L0069GRE. XAC was used as ref compound
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