Adenosine A2AR fluorescent probe
Tag-lite Adenosine A2A receptor fluorescent probe
A2AR expressing cells labeled with Terbium HTRF®
Tag-lite cells transiently expressing the A2A 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
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Transient expression
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Ready for binding
Overview
One of four adenosine receptors found in the brain, A2A is a GPCR. A2A receptor is thought to play a role in acute and chronic neurodegenerative disorders such as cerebral ischemia or Parkinson's disease. A2A may also be involved in seizures and sleep disorders.
Cells expressing the A2A 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 A2A 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 A2A labeled cells and their matching fluorescent ligand (L0058RED). ZM-241385 was used as reference ligand. Results may vary from one HTRF® compatible reader to another.
Compound profiling on Adenosine A1, A2A, A2B and A3 receptors
Adenosine receptors designated by four subtypes A1, A2A, A2B and A3 were shown to be promising pharmacological targets for the development of therapeutic compounds for various therapeutic areas such as cancer, neurodegenerative disease and COPD/Asthma. Thus, a wide range of compounds, and particularly antagonists have been largely studied.
The Tag-Lite® binding assay platform offers the complete adenosine receptors subset A1, A2A, A2B and A3 to characterize compounds in term of binding affinity (Ki) and selectivity profile.
The set of compounds have been profiled on A1, A2A, A2B and A3 receptors using their respective fluorescent ligands L0067RED, L0058RED, L0068RED and L0069GRE.The selectivity profiles of several compounds involved in different therapeutic areas were assessed with this Tag-Lite platform.
The table below summarizes the resulting Ki obtained for each receptor subtype for the full panel of compounds. The results are confirming the expected selectivity and the range of affinities for these reference compounds.
| Compound | Pharmacological class | Compounds affinities (Ki - nM) for the different adenosine receptor subtypes | |||
| A1 | A2A | A2B | A3 | ||
| XAC | Non selective antagonist | 12 | 15 | 20 | 50 |
| Theophylline | Non selective antagonist | 4000 | 5000 | 18000 | 140000 |
| DPCPX | A1 antagonist | 7.6 | 170 | 145 | 800 |
| Vipadenant | A2A antagonist | 70 | 0.9 | 55 | 20000 |
| Ciforadenant (CPI-444) | A2A antagonist | 150 | 3.4 | 660 | 1600 |
| Preladenant (SCH 420814) | A2A antagonist | > 20 000 | 5 | > 20 000 | >20 000 |
| Istradefylline (KW-6002) | A2A antagonist | >10 000 | 40 | >10 000 | >10 000 |
| ZM241385 | A2A antagonist | 260 | 0.7 | 40 | 750 |
| AZD4635 | A2A antagonist | 900 | 15 | 660 | > 10 000 |
| MRS1706 | A2B inverse agonist | 150 | 325 | 30 | > 300 |
| MRS1220 | A3 antagonist | 325 | 37 | > 500 | 1.6 |
The graphs show the pattern of the dose-response curves obtained for five compounds displaying different selectivity profile.
Evaluation of a Tag-lite binding assay for a class B receptor
In collaboration with Boehringer Ingelheim - Scientific Presentations
Ultra HTS at Bayer: use of IP-One and Tag-lite assays in GPCR drug discovery
In collaboration with Bayer - Scientific Presentations
The easy way to approach biology without radioactivity - Flyers
Challenge the limits of binding kinetics studies - Application Notes
HTRF, a powerful assay platform for Immune Checkpoint Studies
Binding Assays to measure therapeutic antitumor antibodies - Posters
Understanding GPCRs is the key to improved DD
In collaboration with GEN - Guides
Tag-lite, the easiest solution for studying GPCR-ligands interactions
The gold standard technology for receptor binding studies - Videos
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
Kinetic adenosine receptor family binding assays
Case study about kinetic binding characterization provided by the combination of Cisbio and Excellerate bioscience expertise. - Application Notes
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Webinar | Association Rate Constant: The Unsung Hero of Drug Discovery Kinetics
Available On-demand - Videos
Easy method for kinetic binding determination
How to revolutionize your kinetic binding demonstration with HTRF kinase binding platform assays - Application Notes
A comprehensive overview of pharmacology's main ligands - Guides
Product Insert A2A LabCell / C1TT1A2A
C1TT1A2A - Product Insert
GPCRs: the pathway to Discovery
Complete solutions for GPCR Drug Discovery - Brochures
Safety Data Sheet (DEU) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Safety Data Sheet (ELL) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Safety Data Sheet (FRA-FR) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Safety Data Sheet (ITA) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Safety Data Sheet (SPA) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Safety Data Sheet (ENG-GB) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Safety Data Sheet (ENG-US) A2A LabCell / C1TT1A2A
C1TT1A2A - Safety Data Sheet
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.
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