Adenosine-to-inosine (A-to-I) RNA editing, a widespread form of RNA modification, involves the conversion of adenosine to inosine in RNA molecules by the action of adenosine deaminases acting on RNA (ADARs). This modification has been thrust into the spotlight due to its substantial impact on biological and pathological processes. It achieves this by altering the structure and function of a variety of RNA molecules, including double-stranded RNA, transfer RNA, microRNA, and viral RNA.

Mounting evidence indicates that A-to-I editing is pivotal in the pathogenesis of numerous diseases, ranging from cancer and autoimmune disorders to cardiovascular diseases, neurological and neurodegenerative conditions, as well as metabolic disorders. Furthermore, the editing of double-stranded RNA (dsRNA) through A-to-I modification also influences other RNA modifications, such as N6-methyladenosine (m6A) and 5-methylcytosine (m5C), thereby reciprocally affecting the development of diseases caused by these modifications.

The RayBio^® A-to-I ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of A-to-I in RNA, serum, plasma and cell medium samples collected from various species. The quantity of A-to-I is determined by a known inosine standard curve. This kit has a high sensitivity of approximately 0.111 ng/mL. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

• Microplate reader capable of measuring absorbance at 450 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1-liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• GraphPad Prism or SigmaPlot software (or other software which can perform four-parameter logistic regression models)
• Tubes to prepare standard or sample dilutions
• Orbital shaker
• Aluminum foil
• Plastic wrap

1. Prepare all reagents, samples and standards as instructed.
2. Add 50 μL standards or samples to appropriate well. Incubate for 10 minutes at room temperature.
3. Add 50 μL anti-inosine to appropriate well. Incubate 1 hour at room temperature. And wash 5 times.
4. Add 100 μL prepared HRP-Secondary antibody solution. Incubate 1 hour at room temperature. And wash 5 times.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 2-10 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

The entire kit may be stored at -20°C to -80°C for up to 1 year from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.