Chemokine CXCR4R expressing cells labeled with Terbium

Tag-lite cells transiently expressing the Chemokine CXCR4 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 Chemokine CXCR4 receptor labeled with Terbium for use in receptor binding applications. The cells are provided ready to use.
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Overview

CXCR4 is a receptor for the C-X-C chemokine CXCL12/SDF-1. Compelling evidences suggest that CXCR4 is nvolved 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. Cells expressing the CXCR4 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 CXCR4 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 CXCR4 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 Chemokine CXCR4 labeled cells and their matching fluorescent ligand (L0012RED). AMD-3100 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
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