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Tag-lite pSNAP-5HT1A Plasmid, 10 µg

pSNAP-5HT1A 5-Hydroxytryptamine 1A image

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.

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Feature	Specification
Application	Receptor-Ligand Binding

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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.

View product information

Product Variant

Unit Size: 1 Item

Part #:

PSNAP5HT1A

List Price

USD 2,098.00

Product information

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

Other Specifications

Application	Receptor-Ligand Binding
Brand	Tag-lite
Detection Modality	HTRF
Product Group	Plasmids
Shipping Conditions	Shipped in Dry Ice
Target Class	GPCR
Technology	TR-FRET
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.

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.

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.

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.