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Glucagon GLP1R stably expressing cells HTRF®

This cell line stably expresses the Glucagon GLP1 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.
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  • Fully functional cell line Fully functional cell line
  • Stable expression Stable expression
This cell line stably expresses the Glucagon GLP1 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.
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Overview

The GLP1 receptor is a GPCR which role is to potentiate the glucose-induced secretion of insulin from pancreatic beta cells, increase insulin expression, inhibit beta-cell apoptosis, promote beta-cell neogenesis.

This HEK293 stable cell line expresses the GLP1 receptor fused to a SNAP-Tag. Labeled with Terbium, the cells can be used in a Tag-lite receptor binding protocol.

Benefits

  • STABLY TRANFECTED
  • HIGH PECENTAGE OF VIABILITY
  • FULLY FUNCTIONAL

Step 1 - Receptor labeling

SNAP-tag®, is a small fusion tag that covalently interacts with specific substrates. SNAP-tag allows specific and 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.

Kd and Ki validation

Examples of data obtained using glucagon GLP1-R expressing stable cells (HEK), labelled with terbium in Tag-lite buffer (LABMED) and their matching fluorescent ligand (L0030RED). Exendin 4 was used as reference ligand. Results may vary from one HTRF® compatible reader to another.

Kd data obtained using glucagon GLP1-R expressing stable cells
Ki data obtained using glucagon GLP1-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

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

HTRF Product Catalog 2020 July update

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

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

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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