Lumit® dsRNA Detection Assays Technical Manual
Instructions for Use of Product(s)
W2041, W2042, W2051, W2052, W2061, W2062, W2071, W2072, W2081, W2082
Literature # TM746
Traditional methods for quantifying dsRNA, such as dot blot or enzyme-linked immunosorbent assay (ELISA), rely on antibody-based detection. However, these methods are often limited by sequence bias, reduced sensitivity and labor intensive protocols.
In contrast, the Lumit® dsRNA Detection Assays offer a homogeneous, bioluminescent approach for dsRNA quantification, eliminating the need for antibodies or wash steps. This assay uses NanoLuc® Binary Technology (NanoBiT), a split luciferase complementation system specifically designed for biomolecular interaction studies. NanoBiT comprises two subunits: Large BiT (LgBiT; 18kDa) and Small BiT (SmBiT; 11 amino acids), both of which are engineered for stability and minimal spontaneous association.
In the Lumit® dsRNA Detection Assay, the sample containing dsRNA is incubated with two dsRNA-binding domains, one labeled with LgBiT and the other with SmBiT. When these binding domains interact with dsRNA that contains standard or chemically modified nucleosides, the LgBiT and SmBiT subunits are brought into proximity, reconstituting the NanoBiT® enzyme and producing luminescence in the presence of the Lumit® substrate. The resulting luminescence is directly proportional to the dsRNA concentration in the sample, enabling rapid and accurate quantification without wash steps.
Summary of Changes
The following changes were made to the 2/26 revision of this document:
1. Added a new protocol for modified standards, Section 5, including Figures 5, 6 and 7.
2. Updated Section 4 to an unmodified standards protocol, including Figures 2, 3 and 4.
3. Added new products to Section 2.
4. Removed the former Section 8, Related Products.
5. Added text to Section 1, Description, noting addition of protocols for unmodified and modified dsRNA Standards.
6. Updated Notes in Section 3, Before You Begin.
7. Added new Causes and Comments to Section 6, Troubleshooting.
Revised 2/26.