CXCR4R stably expressing cells HTRF®

This cell line stably expresses the CXCR4 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.

Fully functional cell line
Stable expression

Download the protocols

This cell line stably expresses the CXCR4 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.

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Overview

CXCR4 is a receptor for the C-X-C chemokine CXCL12/SDF-1. Compelling evidences suggests that CXCR4 is involved in hematopoiesis and in cardiac ventricular septum formation. The receptor also plays an essential role in vascularization of the gastrointestinal tract. CXCR4 acts as a coreceptor for human immunodeficiency virus-1/HIV-1.

This HEK293 stable cell line expresses the CXCR4 receptor fused to a SNAP-Tag. Labeled with Terbium, the cells can be used in a Tag-lite receptor binding protocol.

Benefits

STABLY TRANSFECTED
HIGH PERCENTAGE OF VIABILITY
FULLY FUNCTIONAL

Step 1 - Receptor labeling

SNAP-tag® is a small fusion tag that covalently interacts with specific substrates. It enables specific covalent labeling of any protein of interest. For more details, see the labeling procedure.

Diagram of a chemical reaction between SNAP-tag and its substrate

Watch this video describing how to label cell surface receptors using Tag-lite® technology.

Step 2 - Understand the assay principle

Running a receptor binding assay using Tag-lite is as easy as it can get. 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 a receptor binding assay using the Tag-lite protocol

Step 3 - 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.

Step 4 - 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.

Examples of data obtained using chemokine CXCR4-R expressing stable cells (HEK) labelled with terbium in Tag-lite buffer (LABMED), and their matching fluorescent ligand (L0012RED). AMD-3100 was used as reference ligand. Results may vary from one HTRF® compatible reader to another.

kd determination with cxcr4-R expressing stable cells (HEK)

Ki data obtained using chemokine CXCR4-R expressing stable cells

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

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 - Application Notes

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

Manual CXCR4-R CL (HEK) / C1SU1CXCR4

C1SU1CXCR4 - Product Insert

Combining Kinetic Ligand Binding and 3D Tumor Invasion Technologies to Assess Drug Residence Time and Anti-metastatic Effects of CXCR4 Inhibitors

BioTek - Application Notes

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

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

GPCRs Pharmacology

A comprehensive overview of pharmacology's main ligands - Guides

GPCRs: the pathway to Discovery

Complete solutions for GPCR Drug Discovery - Brochures

Safety Data Sheet (DEU) CXCR4-R CL (HEK) / C1SU1CXCR4