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Tag-lite pSNAP(+) Plasmid, 10 µg
Tag-lite pSNAP(+) Plasmid, 10 µg
Tube Tag-lite GPCR plasmid
| Feature | Specification |
|---|---|
| Application | Receptor-Ligand Binding |
Product variant
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).
Tag-lite pSNAP(+) Plasmid, 10 µg
Tube Tag-lite GPCR plasmid
Tag-lite pSNAP(+) Plasmid, 10 µg
Product information
Overview
Over the past few years, SNAP-Tag technology combined with TR-FRET has paved the road to the development of many non-radioactive, no-wash binding assays. The method is based on transfecting cells using plasmids encoding a SNAP-Tag and subsequently labeling them with Terbium. PSNPPOS is a backbone containing the gene for the SNAP tag, a promoter and MCS (Multiple Cloning Site).
How it works
Step 1 - Plasmid creation
Using standard cloning techniques, insert the GPCR gene of interest into the empty plasmid. Remember that when you are designing a plasmid, the GPCR gene should be kept in the frame.
Step 2 - Plasmid transfection
Use standard transfection techniques (see transient transfection protocol) to transiently express the SNAP-GPCR of interest in your cell line.
Step 3 - 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. Cells are provided unlabeled and need to be labeled with Lumi4-Terbium prior to running a binding assay.
Step 4 - 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 5 - 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.
Step 6 - 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.
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 |
10 µg
|
Video gallery
Tag-lite pSNAP(+) Plasmid, 10 µg
Tag-lite pSNAP(+) Plasmid, 10 µg
Resources
Are you looking for resources, click on the resource type to explore further.
Guide
HTRF solutions, guide to major applications
This guide provides you an overview of HTRF applications in several therapeutic areas.
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