A2AR expressing cells labeled with Terbium

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 Fully functional cell line
  • Transient expression Transient expression
  • Ready for binding Ready for binding
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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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.
Diagram of A2A 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 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.

Representative saturation binding and competitive binding graph
Representative competitive binding graph

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.

CompoundPharmacological classCompounds affinities (Ki - nM) for the different adenosine receptor subtypes


A1A2AA2BA3
XACNon selective antagonist12152050
Theophylline Non selective antagonist4000500018000140000
DPCPXA1 antagonist7.6170145800
VipadenantA2A antagonist700.95520000
Ciforadenant (CPI-444)A2A antagonist1503.46601600
Preladenant (SCH 420814)A2A antagonist> 20 0005> 20 000>20 000
Istradefylline (KW-6002)A2A antagonist>10 00040>10 000>10 000
ZM241385A2A antagonist2600.740750
AZD4635A2A antagonist90015660> 10 000
MRS1706A2B inverse agonist15032530> 300
MRS1220A3 antagonist32537> 5001.6
Adenosine A2A - Assay validation: DPCPX (A1 selective compound) profiling on Adenosine receptors
Adenosine A2A - Assay validation: Preladenant (A2A selective compound) profiling on Adenosine receptors
Adenosine A2A - Assay validation: MRS1706 (A3 selective compound) profiling on Adenosine receptors

The graphs show the pattern of the dose-response curves obtained for five compounds displaying different selectivity profile.

Adenosine A2A - Assay validation: MRS1220 (A3 selective compound) profiling on Adenosine receptors
Adenosine A2A - Assay validation: Theophylline (non selective compound) profiling on Adenosine receptors
Evaluation of a Tag-lite binding assay for a class B receptor

In collaboration with Boehringer Ingelheim - Présentations scientifiaues

Ultra HTS at Bayer: use of IP-One and Tag-lite assays in GPCR drug discovery

In collaboration with Bayer - Présentations scientifiaues

Tag-lite binding assays

The easy way to approach biology without radioactivity - Flyers

Determination of association (kon) and dissociation (koff) rates constants using the Tag-lite platform

Challenge the limits of binding kinetics studies - Notes d'application

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 - Vidéos

Product Insert A2A LabCell / C1TT1A2A

C1TT1A2A Lot 14 - Protocoles

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalogue

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. - Notes d'application

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Easy method for kinetic binding determination

How to revolutionize your kinetic binding demonstration with HTRF kinase binding platform assays - Notes d'application

Safety Data Sheet (DEU) A2A LabCell / C1TT1A2A

C1TT1A2A - Fiches de sécurité

Safety Data Sheet (ELL) A2A LabCell / C1TT1A2A

C1TT1A2A - Fiches de sécurité

Safety Data Sheet (ITA) A2A LabCell / C1TT1A2A

C1TT1A2A - Fiches de sécurité

Safety Data Sheet (SPA) A2A LabCell / C1TT1A2A

C1TT1A2A - Fiches de sécurité

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