Smart detection methods for protein interactions in living cells
In this webinar, we introduce the bioluminescent NanoBiT® and NanoBRET™ technology platforms and their use for studying protein interactions in live cells. NanoLuc® Binary Technology (NanoBiT®) is a structural complementation reporter system based on NanoLuc® luciferase. It is composed of a large subunit called LgBiT (Large BiT; 18kDa) and a small subunit called SmBiT (Small BiT; 1.3 kDa) whose physical interaction reconstitutes the bright NanoBiT® luciferase. Their small size minimizes interference with normal protein function and the low intrinsic affinity mitigates unspecific signal while allowing to monitor interaction dynamics in real-time.
NanoBRET™ represents a powerful alternative to interrogate homo- and heteromeric protein interactions. In this proximity-based assay NanoLuc® is used as BRET energy donor and fluorescently labelled HaloTag® as the energy acceptor. When fused to proteins of interest, this technology enables sensitive, reproducible detection of protein interactions within their cellular context. Furthermore, we present how this live cell format can be adapted to quantitively assess target engagement.
We will highlight the advantages of NanoBiT® and NanoBRET™ and discuss the key steps of their workflows.
You will learn:
• Principle of NanoBiT® and NanoBRET™ technologies
• How to use NanoBiT® for studying protein:protein interaction
• How to use NanoBRET™ for studying protein:protein interaction
• How to use NanoBRET™ for studying target engagement