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HTRF Human STING Binding Kit, 500 Assay Points
HTRF Human STING Binding Kit, 500 Assay Points
Picture of mouse STING binding kit
The human STING WT binding assay is designed to select and characterize compounds that specifically bind human STING protein.
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).
| Feature | Specification |
|---|---|
| Application | Protein-Protein Interaction |
| Sample Volume | 5 µL |
The human STING WT binding assay is designed to select and characterize compounds that specifically bind human STING protein.
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).
Product Variants
Unit Size: 500 Assay Points
Part #:
64BDSTGPEG
List Price
USD 2,091.00
Unit Size: 10,000 Assay Points
Part #:
64BDSTGPEH
List Price
USD 24,550.00
HTRF Human STING Binding Kit, 500 Assay Points
Picture of mouse STING binding kit
HTRF Human STING Binding Kit, 500 Assay Points
Product information
Overview
A fast and easy way to identify new binders to human STING WT (R232).
Stimulator of interferon genes (STING), also known as transmembrane protein 173 (TMEM173), is a protein playing a major role in innate immunity. Upon intracellular cytosolic DNA release from pathogens such as viruses and bacteria, 2’-3’cGAMP binds to STING protein and triggers the secretion of type 1 interferon. Identifying new STING ligands is therefore a way to control immunity and fight autoinflammatory diseases.
Specifications
| Application |
Protein-Protein Interaction
|
|---|---|
| Brand |
HTRF
|
| Detection Modality |
HTRF
|
| Product Group |
Kit
|
| Sample Volume |
5 µL
|
| Shipping Conditions |
Shipped in Dry Ice
|
| 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 STING Binding Kit, 500 Assay Points
HTRF Human STING Binding Kit, 500 Assay Points
How it works
Assay principle
The HTRF STING binding assay is a competitive assay format which uses d2-labeled STING ligand, a 6His tagged human STING protein, and an anti 6His Cryptate-labeled antibody. Your compound competes with the STING ligand-d2 and thereby prevents FRET from occurring.
Assay protocol
The Human STING 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 STING protein is then added followed by the dispensing of the HTRF reagents: the anti 6His antibody labeled with Terbium cryptate and the STING ligand labeled with d2. The reagents labelled 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 details
| STING ligand-d2 - Kd | 15 nM |
|---|---|
| STING ligand-d2 - Concentration | 15 nM |
| 2’3’ cGAMP Standard - IC50 | 5 nM |
| 2’3’ cGAMP Standard - Ki | 2.5 nM |
| Signal to Background | 12.4 |
Assay validation
Compound screening
Various compounds known as STING ligands were added to the assay. 2’-3’ cGAMP (assay standard), cyclic di-AMP, and cyclic di-GMP display the right potency in good correlation with the literature. DMXAA, a non-specific compound, also well-known mouse STING binder, has no effect on the assay, confirming the human specificity of the kit.
Simplified pathway
STING simplified pathway
STING, for STimulator of INterferon Genes, is a cytoplasmic homodimeric protein localized in the endoplasmic reticulum which plays an essential role in innate immunity. Upon pathogen infection or mitochrondrial shrinking during apoptosis, floating dsDNAs bind and activate a DNA sensor, the cyclic GMP-AMP synthase (cGAS). Activated cGAS leads to the production of 2’-3’cGAMP, a cyclic dinucleotide, which then binds to STING proteins. In turn, phosphorylated STING interacts with TANK-binding-kinase-I (TBK1), leading to the recruitment and activation of the active interferon regulatory factor (IRF3) dimer. Nuclear translocation of the IRF3 dimer leads to the transcription of genes encoding IFN-α/β. In addition, the STING pathway controls NF-κB dependent inflammatory cytokine expression. As a negative feedback loop, the DNA-stimulated cGAS-STING-TBK1 pathway also triggers STING protein degradation through p62 SQSTM1 associated autophagy to switch off IFNb production.
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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