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HTRF Tag-Lite pSNAP-β2 Adrenergic Receptor Plasmid

The Tag-lite Beta2 adrenoplasmid is used to transiently or stably transfect cells for the purpose of developing a Beta2 adrenoreceptor binding assay.

For research use only. Not for use in diagnostic procedures.

Part number: PSNAPBD2
List price: USD 2,098.00
Your price:
USD 2,098.00
USD 2,098.00 /each

Overview

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 Beta2 adreno receptor.

Specifications

Assay Points
1
Assay Target Type
Plasmids
Assay Technology
HTRF
Brand
HTRF
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

Video gallery

How it works

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-chemical-reaction
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-psnapbd
how-it-works-psnap-beta2-adrenoreceptor
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-psnapbd
how-it-works-psnap-beta2-adrenoreceptor

 

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.

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SDS, COAs, Manuals and more

Are you looking for technical documents for this product. We have housed them in a dedicated section., click on the links below to explore.

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