Vasopressin V2R stably expressing cells HTRF®

This cell line stably expresses the Vasopressin V2 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 Vasopressin V2 receptor fused to a SNAP-Tag, and can be used in a Tag-lite application.
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Overview

There are three subtypes of vasopressin receptor: V1A, V1B, and V2. The V2 receptor is mainly found in the kidney and is important in free water reabsorption.

This HEK293 stable cell line expresses the V2 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.

Kd and Ki validation

Examples of data obtained using vasopressin V2-R expressing stable cells (HEK) labelled with terbium in Tag-lite buffer (LABMED), and their matching fluorescent ligand (L0063RED). Vasopressin was used as reference ligand. Results may vary from one HTRF® compatible reader to another.

Kd data obtained using vasopressin V2-R expressing stable cells
Ki Data obtained using vasopressin V2-R expressing stable cells

Evaluation of a Tag-lite binding assay for a class B receptor

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Tag-lite binding assays

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The gold standard technology for receptor binding studies - Videos

HTRF Product Catalog 2020 July update

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A guide to Homogeneous Time Resolved Fluorescence

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Get the brochure about technology comparison. - Brochures

Easy method for kinetic binding determination

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