AffiELISA® RIG-I dsRNA ELISA Kit
AffiELISA® RIG-I dsRNA ELISA Kit is a sensitive, high-throughput assay designed to quantify double-stranded RNA (dsRNA) by leveraging the ATPase activity of RIG-I. Upon binding to dsRNA, RIG-I hydrolyzes ATP into ADP and inorganic phosphate. This kit specifically detects ADP production as a readout, providing a functional and quantitative assessment of dsRNA levels in test samples.
AffiELISA® RIG-I dsRNA ELISA Kit is an advanced assay designed to directly evaluate immune responses through the activation of the RIG-I receptor by double-stranded RNA (dsRNA). This kit offers a complementary approach to traditional ELISA methods, enabling a more comprehensive analysis of immune activation.
With a high sensitivity for detecting dsRNA in various sample types, the kit provides valuable insights into innate immunity and viral detection mechanisms.
Intended Use:
AffiELISA® RIG-I dsRNA ELISA Kit is designed for the quantitative detection of double-stranded RNA (dsRNA) and the assessment of RIG-I receptor activation in various biological samples.
Kit components:
|
Components |
Description |
| RIG-I Enzyme | Purified RIG-I protein, which specifically binds and is activated by dsRNA. |
| ATP | Adenosine triphosphate, the substrate for RIG-I's ATPase activity. |
| RIG-I Reaction Buffer | A buffer solution optimized for RIG-I activity and stability. |
| Reagent 1 | A solution designed to deplete residual ATP after the RIG-I reaction, likely containing enzymes like hexokinase and glucose to consume ATP. |
| Reagent 2 | A solution containing enzymes and substrates to convert ADP back to ATP, along with the luciferase/luciferin system for luminescence detection, likely including pyruvate kinase, phosphoenolpyruvate, luciferase, and luciferin. |
| dsRNA Standards | A set of solutions containing known concentrations of dsRNA, used to generate a standard curve. |
Materials Required But Not Supplied:
- Microplate reader capable of reading absorbance at D450/650 nm.
Procedure workflow:
1. RIG-I Reaction:
- Combine RIG-I enzyme, ATP, and sample. Incubate (37°C, 60 min) for RIG-I-mediated ATP hydrolysis to ADP, driven by dsRNA presence.
2. ATP Removal:
- Add Reagent 1 (ATP depletion) and incubate (RT, 60 min) to eliminate residual ATP.
3. Luminescence Generation:
- Add Reagent 2 (ADP-to-ATP conversion, luciferase/luciferin). Incubate (RT, 40 min) for luminescence production, proportional to initial ADP.
4. Luminescence Measurement:
- Read luminescence using a plate reader.
5. dsRNA Quantification:
- Determine dsRNA concentration by comparing luminescence to a dsRNA standard curve.