NanoBiT PPI Control Pair (FKBP, FRB)

NanoLuc Binary Technology (NanoBiT) is a two-subunit system based on NanoLuc luciferase that can be applied to the intracellular detection of protein:protein interactions (PPIs) in live cells. The NanoBiT system is composed of two small subunits, Large BiT (LgBiT; 18kDa) and Small BiT (SmBiT; 11 amino acid peptide), that are expressed as fusions to target proteins of interest. The LgBiT and SmBiT subunits have been independently optimized for stability and minimal self-association. Interaction of the target proteins facilitates subunit complementation to give a bright, luminescent enzyme. The NanoBiT PPI Starter Systems provide the vectors required to create the LgBiT and SmBiT protein fusions, a PRKACA:PRKAR2A constitutively interacting positive control pair and a negative control vector. Starter systems also include the Nano-Glo Live Cell Assay System, a single-addition, non-lytic detection reagent used for monitoring NanoBiT luminescence in living cells. The reagent is prepared by diluting the Nano-Glo Live Cell Substrate with the Nano-Glo LCS Dilution Buffer to make the Nano-Glo Live Cell Reagent. Both substrate and buffer solutions are optimized to provide enhanced stability and reduce autoluminescence in the presence or absence of serum, increasing the sensitivity for detection of low levels of NanoBiT luminescence. The FKBP:FRB pair is provided separately as an inducible positive control. Expression is driven by HSV-TK promoter, providing constitutive, low-level expression in mammalian cells. Using the NanoBiT MCS Starter System, you can generate N- and C-terminal LgBiT and SmBiT fusions to proteins of interest using traditional cloning with a multiple cloning site (MCS). Using the NanoBiT Flexi Starter System, you can generate N- and C-terminal LgBiT and SmBiT fusions using the Flexi Vector Cloning System, a directional cloning method based on two rare-cutting restriction enzymes, SgfI and PmeI, that provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between Flexi Vectors without the need to resequence. Utilize Find My Gene to obtain your ORF clones already in Flexi format for simple creation of fusions. To learn more, visit: www.promega.com/NanoBiT