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Caspase-3 activity biosensor measured using BRET

BRET-based Biosensors for Specific Intracellular Event

Biosensors based on intramolecular changes in Bioluminescence Resonance Energy Transfer (BRET) have been used to interrogate a number of different signaling events inside and outside of the cell¹. These sensors rely on the energy from luciferase acting as a BRET donor, and this energy is transferred to a fluorescent BRET acceptor based on the proximity of the two partners. NanoLuc® Luciferase has been used to generate these biosensors and offers unique advantages due to its bright light output and small size.

¹ Salahpour, A. et al. (2012) BRET biosensors to study GPCR biology, pharmacology, and signaling. Frontiers in Endocrinol. 3, 105.

Step 1: Construct Your Biosensor Molecule

Choose a NanoLuc® Vector and a HaloTag® Vector to construct your biosensor of interest. Or, consult with our Custom Assay Services team to explore the creation of a custom solution.

We offer a range of vectors designed for simple generation of NanoLuc® luciferase N- or C-terminal fusion proteins with CMV-driven expression and additional NanoLuc® Vectors in a variety of configurations for additional cloning options.

View NanoLuc® Fusion Vectors View Additional NanoLuc® Vectors Request NanoLuc® Vector Sample

HaloTag® is a convenient protein fusion tag that binds the NanoBRET™ fluorescent ligand, which serves as the BRET acceptor. Numerous options are available to design HaloTag® N- or C-terminal fusion proteins in an expression vector. Learn more about HaloTag® Technology here.

View HaloTag® N-Terminal Vectors View HaloTag® C-Terminal Vectors

Step 2: Detect BioSensor Activity

The NanoBRET™ Nano-Glo® Detection System provides 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. When paired with HaloTag® Protein Labeling Technology, NanoBRET™ 618 has advantages in Stokes shift and signal:background ratio over other available BRET acceptors. NanoBRET™ Detection System is available from Promega Custom Assay Services.

Ordering Information
GloMax Discover Instrument

The NanoBRET™ assay is detected with a microplate luminometer, such as the GloMax® Discover. Learn more about GloMax® Discover here.

Explore NanoLuc® Luciferase

Discover NanoLuc® Luciferase

The small size and bright luminescence of NanoLuc® Luciferase bring exquisite sensitivity to reporter assays and new possibilities to diverse bioluminescence applications.

Discover NanoLuc® Luciferase
Protein:Protein Interactions

New ways to understand protein interactions, identify the partners involved, and decipher their biological significance.

Protein:Protein Interactions
Target Engagement

Quantify the interaction between a molecule and a protein in real time, live cell assays.

Target Engagement
Protein Stability

Monitor changes in intracellular protein abundance with NanoLuc® fusion proteins.

Protein Stability
Gene Regulation & Cell Signaling

Greater sensitivity and versatility for traditional gene regulation assays.

Gene Regulation & Cell Signaling

Sensitive, bright bioluminescent imaging in cells and whole animals.

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