Protein:Protein Interaction (PPI) Assays use NanoLuc Luciferase and HaloTag protein fused to target proteins of interest to enable sensitive, reproducible detection of protein interactions in the natural cellular environment.

• BRET-Based Assays to Detect Protein:Protein Interactions in Live Cells
• Study induction and inhibition of protein interactions using full-length proteins expressed at physiologically relevant levels
• Starter Systems provide fusion vectors, positive controls and detection reagents for building PPI assays
• Choice of multicloning site (MCS) or Flexi(R) vector-based cloning options

NanoBRET technology is an improved bioluminescence resonance energy transfer (BRET)-based technology that uses NanoLuc luciferase as the BRET energy donor and HaloTag protein labeled with the NanoBRET 618 fluorescent ligand as the energy acceptor to measure the interaction of two binding partners in live cells. NanoBRET Protein:Protein Interaction (PPI) Assays use NanoLuc Luciferase and HaloTag protein fused to target proteins of interest to enable 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. For more details on using NanoBRET technology for protein:protein interaction studies visit the NanoBRET Technology for Protein Interactions page. The NanoBRET PPI Starter Systems provide the vectors required to create NanoLuc Luciferase and HaloTag protein fusions to target proteins of interest, the NanoBRET PPI Positive Control Pair (p53,MDM2) and the NanoBRET Nano-Glo Detection System, which contains the NanoBRET Nano-Glo Substrate used by NanoLuc Luciferase to generate the donor signal and the HaloTag NanoBRET 618 Ligand for the fluorescent energy acceptor. MCS Starter System: Generate N- and C-terminal NanoLuc Luciferase and HaloTag protein fusions to target proteins using traditional cloning with a multiple cloning site (MCS). Flexi Starter System: Generate N- and C-terminal NanoLuc Luciferase and HaloTag protein 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 as HaloTag fusions.