NanoLuc® Binary Technology, or NanoBiT, is a new complementation reporter system based on NanoLuc® Luciferase for tagging proteins and monitoring protein interactions within living cells.
NanoLuc® Luciferase is a small protein that catalyzes a bright luminescent reaction.
To create a complementation system, NanoLuc® Luciferase was separated into two subunits that have very little enzymatic activity by themselves.
The LgBiT has been engineered to stabilize its structure.
The SmBiT was selected from a peptide library because of its low affinity for the LgBiT.
The low affinity SmBiT needs to be brought into close proximity to the LgBiT to facilitate re-association and restore enzymatic activity making it perfect for studying protein:protein interactions.
Because the subunits have low affinity for each other, their interaction is driven by the behavior of their fusion partners.
Their interaction is reversible and both association and dissociation of the target proteins can be monitored.
There is no lag time in the loss of luminescence so detection is immediate allowing for accurate temporal dynamics.
The small size of both bits reduce problems from steric hindrance, and the tiny size of the SmBiT opens other possibilities for its use
like in the study of viral proteins where encapsulation limits the size of gene fusions.
The NanoBiT™ Assay uses small tags that minimize steric effects,
has a bright signal providing high sensitivity even at low expression levels
and allows you to accurately observe association and dissociation of target proteins.
These properties of the NanoBiT™ Assay have been optimized to cause minimal interference with the proteins being studied giving you more accurate results.