Product description

Retinoic acid-inducible gene I (RIG-I) functions as a cytosolic pattern recognition receptor that initiates innate antiviral immunity by detecting exogenous viral RNAs. Structurally, RIG-I is comprised of three main functional domains – Caspase Activation and Recruitment Domain (CARD) for signal transduction, DExD/ H-box RNA helicase domain, and C-terminal domain for RNA binding. The RNA helicase domain is associated with RNA-dependent ATP hydrolysis activities, which is activated upon binding to double-stranded RNA (dsRNA), leading to the hydrolysis of ATP into ADP and inorganic phosphate (Pi). This kit quantifies dsRNA content in samples by measuring ADP production during the reaction between RIG-I and the sample. The assay is performed in a multi-well plate and can detect reactions in volumes as low as 5 μL. 

The detection involves three steps:

1. RIG-I Reaction: Mix RIG-I enzyme, ATP, and the sample, followed by incubation. 

2. ATP Depletion: Add an equal volume of Reagent 1 to deplete residual ATP. 

3. ADP Detection: Add Reagent 2 to convert ADP back to ATP, which is then quantified using a luciferase/luciferin reaction. The luminescent signal is measured with a plate reader. dsRNA content is calculated based on a standard curve.

Kit Components

Core Principle

When RIG-I binds to dsRNA, its ATPase activity is triggered, hydrolyzing ATP into ADP and Pi. This product evaluates the immunogenicity of dsRNA in samples by measuring the amount of ADP generated during the RIG-I/sample reaction.

Product Advantages

· Direct Immunogenicity Assessment via Receptor Activation In Vitro
A breakthrough from traditional detection methods, directly evaluating sample immunogenicity based on receptor activation levels.

· High Sensitivity for Short-chain RNA
Highly sensitive detection of short dsRNA sequences, superior performance for short fragments.

· Complementary Evaluation with ELISA
Works in conjunction with ELISA, offering a comprehensive assessment of sample immunogenicity.

Data Presentation

Specific Recognition

Recognition of Short dsRNA

RIG-I

ELISA antibody

RIG-I shows superior recognition of short dsRNA compared to antigen-antibody recognition in ELISA.

Comparison of Immunogenicity Detection Results Using Different Methods