RIG-1 dsRNA Detection Kit HYJ522

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.