NanoLuc® Binary Technology (NanoBiT) is a structural complementation reporter designed for protein:protein interaction (PPI) studies. NanoBiT has two small subunits optimized for stability, low self-affinity and bright luminescence. If two proteins of interest are tagged with these subunits and then interact, the subunits come together to form an active enzyme. The proportional signal-to-activity and plate-based format make NanoBiT ideal for PPI studies, including high-throughput screening.
Design PPI assays that more accurately reflect natural cellular biology. Small subunit size (LgBiT, 18kDa and SmBiT, 11 amino acids) minimizes interference with normal protein function, and the bright signal accommodates low, native expression levels. NanoBiT® subunit interactions are reversible and can detect rapidly associating and dissociating proteins. Use NanoBiT in live cells for accurate, real-time monitoring of protein dynamics within a natural cellular context.
Read more about NanoBiT® technology in Dixon et al., 2015:
NanoLuc Complementation Reporter Optimized for Accurate Measurement of Protein Interactions in Cells
Exciting possibilities for exploring protein interactions in live cells
The optimized NanoBiT® reporters open up a world of options for exploring natural biology. See how NanoBiT® technology is inspiring scientists to take their research a little BiT further in the following videos.
Watch Protein Interactions in Real Time Inside Cells
HeLa cells were transfected with FKBP-SmBiT and FRB-LgBiT fusion constructs. Following overnight incubation, substrate was added and samples imaged on an LV200 microscope. After baseline read for approximately 30 seconds, 500nM rapamycin was added to induce FKBP:FRB interaction and images were recorded (imaging cycle = 0.5 seconds).
Monitor Induction of Protein Interactions
Monitor Inhibition of Protein Interactions
Understanding the NanoBiT® PPI System
NanoBiT is a structural reporter that uses two small complementary subunits as fusion partners with proteins of interest. When those proteins interact, the two subunits of NanoBiT are brought together to form an active enzyme.
Exploring PPI with SmBiT: LgBiT Fusions
Improved Brightness and Accuracy Over Split Firefly Luciferase.
A Dynamic, Reversible System
Brightness Enables Single Copy Integration.