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  • Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points

Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points

Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
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Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Video Play Button
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells)
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
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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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Feature Specification
Application Receptor-Ligand 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.

View product information
Product variant
Unit Size: 200 assay points
Part #:
C1TT1A3
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).
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Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Video Play Button
Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Video Play Button
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells)
Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Diagram of Adenosine A3 receptor labeled with Tb (transiently expressing cells) image
Video Play Button
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Product information

  • Overview
  • How it works
  • Assay validation
  • Specifications
  • Video gallery

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.

How it works

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.

gpcr-how-it-works-assay-principle-a3-receptor-c1tt1a3
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.

gpcr-how-it-works-saturation-binding-kd-c1tt1a3
how-it-works-adenosine-a3-receptor-labeled-cells
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.

gpcr-how-it-works-competitive-binding-ki-c1tt1a3
how-it-works-adenosine-a3-receptor-labeled-cells

 

Assay validation

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.

assay-validation-adenosine-a3-labeled-cells
assay-validation-adenosine-a3-labeled-cells
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 14000
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 >20000 5 >20000 >20000
Istradefylline (KW-6002) A2A antagonist >10000 40 >10000 >10000
ZM241385 A2A antagonist 260 0.7 40 750
AZD4635 A2A antagonist 900 15 660 >10000
MRS1706 A2B inverse agonist 150 325 30 >300
MRS1220 A3 antagonist 325 37 >500 1.6


 

assay-validation-adenosine-a3-labeled-cells
assay-validation-adenosine-a3-labeled-cells

 

assay-validation-adenosine-a3-labeled-cells

 

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

assay-validation-adenosine-a3-labeled-cells

 

assay-validation-adenosine-a3-labeled-cells

 

 

Specifications

Application
Receptor-Ligand Binding
Brand
Tag-lite
Detection Modality
HTRF
Product Group
Cells
Shipping Conditions
Shipped in Dry Ice
Target Class
GPCR
Technology
TR-FRET
Therapeutic Area
Cardiovascular
Infectious Diseases
Inflammation
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
Rare Diseases
Unit Size
200 assay points

Video gallery

Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points
Tag-lite Adenosine A3 receptor Frozen & Labeled Cells, 200 Assay Points

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1-4 of 4 Resources
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Technical Note
A simple method for preparing GPCR membrane model extracts from stable cell lines for use with the HTRF GTP Gi binding assay

G-protein coupled receptors (GPCRs) are crucial transmembrane proteins involved in cellular signal transduction.

This technical...

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Brochure
HTRF assays and reagents product list

Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a...

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Guide Icon
Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

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Flyer
Tag-lite: the binding receptor assay portfolio addressing the adenosine receptor family

Tag-lite™ is a cell-based, no-wash alternative to traditional radioisotope assays for studying GPCR-ligand interaction.

Adenosine...

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