Products from BPS Bioscience require a minimum order value above 400€Applications: Study RNA-binding kinetics and use in high throughput screening (HTS) applications.
Background: ADAR1 (adenosine deaminase, RNA-specific 1) performs adenosine to inosine base editing in RNA, particularly targeting adenosines located within a specific stem-loop motif structure. It is proposed that ADARs evolved to provide additional diversity to the transcriptome and while the majority of ADAR editing events occur in non-coding RNAs, some, including the canonical GluA2 editing site, alter the amino acid sequence of coding proteins. ADAR1 plays a role in innate immunity by mitigating interferon signaling. Dysfunction of ADAR1 results in autoimmune disorders, and impacts cancer cell growth and proliferation as well as tumor response to immunotherapy. Since ADAR recognizes double-stranded RNA, it also functions to suppress or modify RNA viruses. Thus, it is implicated in viral evolution and in the emergence of viral variants such as SARS-CoV-2 variants.
Contraindications: This kit is compatible with up to 1% final DMSO concentration.
Description: The ADAR1:RNA TR-FRET Assay Kit is designed to measure the binding of ADAR1 (adenosine deaminase, RNA-specific 1) to RNA for screening and profiling applications using TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer). It utilizes Terbium-labeled donor and dye-labeled acceptor to complete the TR-FRET pairing. The ADAR1:RNA TR-FRET Assay Kit comes in a convenient 384-well format, with enough purified ADAR1, Tb-Labeled Donor and Dye-Labeled Acceptor, RNase inhibitor, ADAR1 substrate and assay buffer for 384 reactions. The assay also includes a Competitor RNA as control.
Figure 1: ADAR1 mechanism of action on RNA molecules.
Figure 2: Assay principle.
The Terbium-labeled anti-FLAG antibody donor binds to FLAG-tagged ADAR1, while the dye-labeled streptavidin acceptor binds to the biotinylated RNA substrate. When ADAR1 forms a complex with its RNA substrate TR-FRET occurs and can be measured using a TR-FRET fluorescence plate reader. TR-FRET signal is proportional to RNA binding.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed
Uniprot: P55265
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Keegan L. P., et al., 2023 Acc Chem Res 56 (22): 3165-3174.
Mendoza H. G., et al., 2023 Biochemistry 62 (8): 1367-1387.
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