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HTRF Tag-Lite pSNAP-D2 Dopamine Receptor type 2 Plasmid

The Tag-lite D2 Dopamine type 2 plasmid is used to transiently or stably transfect cells for the purpose of developing a D2 Dopamine type 2receptor binding assay.

For research use only. Not for use in diagnostic procedures.
Part Number: PSNAPD2
Part Number: PSNAPD2
  • Over past few years, SNAP-Tag technology combined with TR-FRET has paved way development of many non-radioactive, no-wash, binding assays. method is based on transfecting cells using plasmids encoding a SNAP-Tag and subsequently labeling m with Terbium. Revvity offers a large collection of such plasmids. All GPCR genes are cloned in an expression vector directly downstream from a CMV promoter, and are provided ready protein expression and labeling.

    All information on this page pertains to Tag-lite plasmid cloned with D2 Dopamine type 2 receptor.

  • Assay Points
    1
    Assay Target Type
    Plasmids
    Assay Technology
    HTRF
    Brand
    Tag-Lite
    Quantity
    1
    Shelf Life
    3650.0 Day(s)
    Shipping Conditions
    Shipped in Dry Ice
    Therapeutic Area
    Cardiovascular
    Infectious Diseases
    Metabolism/Diabetes
    NASH/Fibrosis
    Neuroscience
    Oncology & Inflammation
    Rare Diseases
    Unit Size
    1 Item
  • Step 1 - Plasmid transfection

    Use standard transfection techniques (refer to transient transfection protocol) to transiently express the SNAP-GPCR of interest in your cell line.

    Step 2 - Receptor labeling

    SNAP-tag® is a small fusion tag that covalently interacts with specific substrates. It allows specific and covalent labeling of any protein of interest (refer to labeling procedure). Cells are provided unlabeled and need to be labeled with Lumi4-Terbium prior to running a binding assay. Labeling reagents are available from the Revvity catalog in 4 different sizes.

    gpcr-how-it-works-step-2-receptor-labeling
    Step 3 - 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.

    gpcr-how-it-works-step-3-understand-the-assay-principle-receptor-binding
    Step 4- 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.

    gpcr-how-it-works-step-4-saturation-binding-kd-psnapd
    how-it-works-psnap-d2-dopamine-receptor-type-
    Step 5 - 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.

    gpcr-how-it-works-step-5-competitive-binding-ki-psnapd
    how-it-works-psnap-d2-dopamine-receptor-type

     

Resources

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Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.