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HTRF Tag-Lite pSNAP-5HT1A Receptor Plasmid

The Tag-lite 5HT1A 5-Hydroxytryptamine 1A plasmid is used to transiently or stably transfect cells for the purpose of developing a 5HT1A 5-Hydroxytryptamine 1Areceptor binding assay.

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

Part number: PSNAP5HT1A
List price: USD 2,098.00
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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 5HT1A 5-Hydroxytryptamine 1A receptor.

Specifications

Assay Points
1
Assay Technology
HTRF
Brand
Tag-Lite
Product Group
Plasmids
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. SNAP-tag 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
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-psnap5ht1a
how-it-works-psnap-5ht1a-5-hydroxytryptamine
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
how-it-works-psnap-5ht1a

 

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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