Revvity, PSNAPA3, Tag-lite pSNAP-Adenosine A3 receptor Plasmid, 10 µg

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 Adenosine A3 receptor.
Tag-lite assays offer many features and applications over traditional receptor/ligand binding assays:

Non-radioactive
Homogeneous and filtration-free
Ready-to-use kits and reagents for binding assays
Does not alter the receptor pharmacology
Peer-reviewed, validated technology
Thorough kinetic studies with true Kd and Ki values, association (Kon) and dissociation (Koff) rate constants
Screening and profiling of biologics and large molecules
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 enables the 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 in 4 different sizes from the Revvity catalog.

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.

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. Application-Receptor-Ligand Binding
Brand-HTRF
Detection Modality-HTRF
Product Group-Plasmids
Shipping Conditions-Shipped in Dry Ice
Target Class-GPCR
Technology-TR-FRET
Therapeutic Area-Cardiovascular Infectious Diseases Inflammation Metabolism/Diabetes NASH/Fibrosis Neuroscience Oncology & Inflammation Rare Diseases
Unit Size-1 unit