Dopamine D2R stably expressing cells

This cell line stably expresses the Dopamine D2 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.
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  • Fully functional cell line
  • Stable expression
This cell line stably expresses the Dopamine D2 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.
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Overview

Dopaminergic neurons and dopamine receptors are the most numerous in brain. In the brain, dopamine receptors D1 and D2 are GPCR responsible for such crucial processes as voluntary movement, sleep, regulation of food intake, attention, learning, and working memory.

This HEK293 stable cell line expresses the D2 receptor fused to a SNAP-Tag. Labeled with Terbium, the cells can be used in a Tag-lite receptor binding protocol.

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.

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.

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.

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.

Watch this video explaining how to run a competitive binding assay using Tag-lite.

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

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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

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

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