Conventional immunoassays, such as Western blots, sandwich ELISAs or other enzyme immunoassays for studying drug targets, can be time-consuming and produce variable results. Lumit™ Immunoassays offer direct luminescence measurement in cell culture, broad dynamic range, and can be completed in as little as 30 minutes.
Lumit™ Immunoassays are based on NanoLuc® Binary Technology (NanoBiT®). In Lumit™ Immunoassays, antibodies are chemically labeled with the small and large subunits of NanoLuc® Luciferase, known as SmBiT and LgBiT, respectively. In the presence of an analyte, the two antibodies come into close proximity, allowing SmBiT and LgBiT to form an active enzyme and generate a bright luminescence signal.
Advantages over conventional immunoassays, such as ELISAs:
- Simple add-mix-read format: Complete in as little as 30 minutes using a standard luminometer.
- Direct analyte measurement: Detect luminescent signal in the cell culture plate or on medium removed from the cells.
- Broad dynamic range: Reduces the need for sample dilutions.
What Are Immunoassays?
The detection and quantitation of individual proteins is one of the fundamental aspects of proteomics. Immunological-based methods, such as quantitative enzyme-linked immunosorbent assays (ELISA), have been the standard in protein detection for 40 years. The main challenges with interpretation of ELISA data are background noise and assay variability. The well-washing process needs to be as thorough as possible to remove any unbound material and provide optimized assay conditions.
Another popular protein detection method is the Western blot. In this method, proteins are separated by denaturing polyacrylamide gel electrophoresis and transferred to a membrane, such as nitrocellulose or polyvinylidene difluoride (PVDF). After blocking the membrane to reduce nonspecific binding, antibodies targeted to the protein of interest are added. A variety of techniques can then be used to visualize the protein-antibody complexes, including chemiluminescence, colorimetry, immunofluorescence or radioactivity.
No-wash, in solution immunoassays offer considerable advantages over both ELISAs and Western blots. These assays are performed following simple “add-mix-read” protocols with reduced hands-on time. They eliminate the need for multiple washes to separate bound from unbound assay components.