JP
Revvity Sites Globally
Select your location.
*e-commerce not available for this region.
HTRF Human RIG-1 Binding Kit, 500 Assay Points
HTRF Human RIG-1 Binding Kit, 500 Assay Points
Generic HTRF RIG I binding image
The human RIG-I binding assay is designed to select and characterize compounds that specifically bind human RIG-I protein.
| Feature | Specification |
|---|---|
| Application | タンパク質間相互作用 |
| Sample Volume | 5 µL |
The human RIG-I binding assay is designed to select and characterize compounds that specifically bind human RIG-I protein.
Product variants
Unit Size: 500 assay points
Part #:
64BDRIGIPEG
Unit Size: 10,000 assay points
Part #:
64BDRIGIPEH
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).
HTRF Human RIG-1 Binding Kit, 500 Assay Points
Generic HTRF RIG I binding image
HTRF Human RIG-1 Binding Kit, 500 Assay Points
Product information
Overview
A fast and easy way to identify new binders to human RIG-I. The Retinoic acid-Inducible Gene I (RIG-I) is a cytosolic pattern recognition receptor which is known to recognize RNA pathogens in cells. This protein is involved in the innate immune system and induces a type 1 interferon response. The RIG-I protein recognizes infected cells by detecting viral dsRNA. Once activated by a dsRNA, the N-terminus domain of RIG-I binds mitochondrial antiviral signaling proteins, thereby activating the IFN-1 signaling pathway.
HTRF assays offer many advantages over other technologies:
- Homogeneous add-and-read format
- No wash steps
- Low background
- Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
- Stable signal, providing flexibility in time of readout or size of assays
How it works
Assay principle
The HTRF RIG-I binding assay is a competitive assay format which uses a biotinylated 3 phosphate-dsRNA, a streptavidin-d2, a 6His tagged human RIG-I protein, and an anti 6His Cryptate-labeled antibody. The compound being tested competes with the biotinylated 3p-dsRNA, and thereby prevents FRET from occurring.
Assay protocol
The Human RIG-I binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate. The human His-tagged RIG-I protein is then added, followed by the dispensing of the HTRF reagents: The anti 6His antibody labeled with Terbium cryptate and the biotinylated-3p-dsRNA bound to d2-labeled streptavidin. The reagents labeled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
Assay validation
Effect of 5' triphosphate vs blunt-end dsRNA for RIG-I binding
Two different ds-RNAs were tested in parallel in the HTRF RIG-I binding assay. The first ds-RNA displays a triphosphate in 5', the second does not. Serial dilutions of both RNAs were carried out in diluent, and then RIG-I protein was dispensed. HTRF detection reagents were then added to the well for the final detection step. Results were collected after 1 hour of incubation at room temperature. The affinity of the RIG-I protein was increased by the presence of the triphosphate in 5' of the dsRNA, as expected.
Specifications
| Application |
Protein-Protein Interaction
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
HTRF
|
| Detection Modality |
HTRF
|
| Product Group |
Kit
|
| Sample Volume |
5 µL
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Target |
RIG-1
|
| Target Class |
Binding Assay
|
| Target Species |
Human
|
| Technology |
TR-FRET
|
| Therapeutic Area |
Infectious Diseases
Oncology & Inflammation
|
| Unit Size |
500 assay points
|
Video gallery
HTRF Human RIG-1 Binding Kit, 500 Assay Points
HTRF Human RIG-1 Binding Kit, 500 Assay Points
Resources
Are you looking for resources, click on the resource type to explore further.
Brochure
HTRF assays and reagents product list
Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a...
Guide
HTRF solutions, guide to major applications
This guide provides you an overview of HTRF applications in several therapeutic areas.
How can we help you?
We are here to answer your questions.
