Luminescence (incl. BRET)
In life sciences, luminescence detection is an important tool to monitor biological activities. The advantage of luminescent assays mainly is a broad dynamic range with low background. This makes it the ideal tool for the detection of reporter gene assays and all applications requiring high sensitivity.
Luminescence reactions produce light as a result of a chemical or biological reaction, making an excitation source dispensable. In fact, luminescence plate readers require only a reading chamber, a detector (usually a photomultiplier tube) and an optical system bringing the light signal of the sample to the detector. In this optical system, filters or a wavelength monochromator can be employed to select specific luminescent wavelengths or ranges thereof. This helps optimizing the assays by reducing the noise generated by unwanted wavelengths. Moreover, it is useful in the development of new assays or when detecting multiple luminescent signals.
Luminescence reactions can be divided into two categories: glow and flash assays. Glow assays are quite slow, less intense but stable over a longer time. Flash reactions on the contrary are more intense but only for a very short amount of time (seconds). Flash assays typically require the injection of a compound in order to start the reaction and hence can only be performed on microplate readers with reagent injectors.
All luciferase-based gene expression assays, ATP detection-based cell viability/cytotoxicity assays, as well as Bioluminescence Resonance Energy Transfer (BRET) assays used for interaction studies are a few common applications.
Notas de aplicação
- NanoBRET™ assay quantitatively evaluates VEGF binding to the VEGFR2 in real-time in living cells
- CRISPR/Cas9 genome-edited cells express nanoBRET-donor that monitors protein interaction and trafficking
- Real-time assessment of apoptosis and necrosis
- Verifying SPARCL performance on the CLARIOstar equipped for reading at time of injection
- NanoBRET assay for monitoring of ligand binding to GPCRs in live cells, using the CLARIOstar and the PHERAstar FS