What is NanoBRET?

NanoBRET™ Technology enables sensitive, reproducible detection of protein interactions in the natural cellular environment. The use of full-length proteins expressed at low levels enables PPI monitoring and screening studies that reflect true cellular physiology.

What is luciferase reporter assay?

A luciferase reporter assay is a test that investigates whether a protein can activate or repress the expression of a target gene using luciferase as a reporter protein (Carter & Shieh, 2015).

What is NanoBit assay?

The NanoBit ® assay provides a tool for detecting protein-protein interactions in live cells ( Figure 1A). The assay is based on splitting the engineered luminescent protein NanoLuc ® into two separate subunits, the small BiT (SmBiT, 1.3 kDa in size) and the Large BiT (LgBiT, 18 kDa in size).

What is a HiBiT assay?

The HiBiT assay is based on bioluminescence, which is more sensitive than fluorophore-based methods, such as GFP. This allows detection at extremely low levels. Not only is it sensitive, it is also highly quantitative.

How is luciferase assay done?

Cell debris is removed by microcentrifugation and luciferase activity is measured using a luminometer. Some luminometers directly inject the reagents into the cell lysate. Such automation allows the signal to be measured at a precise time following injection, which can increase the consistency of the results.

What are luciferase used for?

Luciferases are widely used in biotechnology, for bioluminescence imaging microscopy and as reporter genes, for many of the same applications as fluorescent proteins.

What is HiBiT?

Scientists at Promega have developed a new way to tag endogenous proteins that offers an alternative to antibody-based assays. The tag is called HiBiT—a small 11 amino acid peptide that binds with high affinity to another larger subunit called LgBiT.

When was luciferase discovered?

While DuBois discovered the reaction between luciferin and luciferase in 1885, it was not until the late 1940s when the luciferase protein was first extracted and purified firefly lanterns by Drs. Green and McElroy. Using this process, they isolated the enzyme and determined its conformational structure.

What is a Hibit tag?

The tag is called HiBiT—a small 11 amino acid peptide that binds with high affinity to another larger subunit called LgBiT. The bound complex has luciferase activity, and will release luminescent signal in the presence of added furimazine substrate. In a recent study published in ACS Chem.

What is endogenous expression?

developing or originating within an organism or part of an organism […]