NanoBRET™ Protein:Protein Interaction System Technical Manual

Instructions for Use of Product(s)
N1661, N1662, N1663, N2583, N2584, N2585, N1811, N1821, N1581, N1641

Literature # TM439

The NanoBRET™ Protein:Protein Interaction (PPI) System is a new configuration and approach to bioluminescence resonance energy transfer (BRET), a proximity-based assay that can detect protein interactions by measuring energy transfer from a bioluminescent protein donor to a fluorescent protein acceptor. The NanoBRET™ assay uses a NanoLuc® fusion protein as the energy donor and a fluorescently labeled HaloTag® fusion protein as the energy acceptor. The optimized blue-shifted NanoLuc® donor paired with the red-shifted HaloTag® acceptor minimizes spectral overlap within the assay for an improved signal:background ratio when calculating the NanoBRET™ ratio. Measure live-cell PPI using an endpoint or kinetic assay.

Summary of Changes
The following changes were made to the 10/23 revision of this document:
1.The y-axis labels for Figures 6, 7 and 8 were updated.
2. A column header for a table in Section 5.A was updated.

Revised 10/23.