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Protein Stability

Protein stability probe using NanoLuc Luciferase

Monitor Protein Dynamics in Vivo

The enhanced brightness and small size of NanoLuc® luciferase make it an ideal protein fusion tag that provides a sensitive and quantitative method for monitoring changes in intracellular protein abundance.

The intracellular level of many proteins is tightly controlled by a balanced rate of synthesis and degradation, and many proteins are maintained at low intracellular abundance by targeting for proteolysis via the ubiquitin/proteasome system. Certain conditions, such as cellular stress, result in increased protein stabilization causing accumulation and subsequent activation of downstream signaling events. Regulation of protein stability is the primary method of regulation for many signaling pathways. The ability to monitor changes in the intracellular protein abundance is valuable for understanding the cellular response to specific treatment or conditions.

Read more:

Measuring Intracellular Protein Lifetime Dynamics Using NanoLuc® Luciferase

NanoLuc® Stability Sensors for HIF1A and NRF2 Proteins

Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.

Panel A. HCT-116 cells were transiently transfected with the HIF1A-NanoLuc fusion vector and Transfection Carrier DNA.

Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.

Panel A. HCT-116 cells were transiently transfected with the HIF1A-NanoLuc fusion vector and Transfection Carrier DNA.

Rapid stabilization and accumulation of NRF2 monitored with NRF2-NanoLuc® fusion protein.

Panel B. HCT-116 cells were transiently transfected with the NRF2-NanoLuc fusion vector and pKEAP1 DNA. Treatments occurred as indicated and NanoLuc® detected using Nano-Glo Luciferase Assay at the indicated time points.

Rapid stabilization and accumulation of NRF2 monitored with NRF2-NanoLuc® fusion protein.

Panel B. HCT-116 cells were transiently transfected with the NRF2-NanoLuc fusion vector and pKEAP1 DNA. Treatments occurred as indicated and NanoLuc® detected using Nano-Glo Luciferase Assay at the indicated time points.

Isogenic Cell Lines with NanoLuc® Fusions

In partnership with Horizon Discovery, a number of reporter cell lines have been created using adenovirus-mediated genome editing to insert NanoLuc® luciferase and create a gene encoding a fusion protein at the endogenous locus of the protein of interest. Learn more about Isogenic Reporter Cell Lines.

Step 1: Choose a Vector to Create a NanoLuc® Fusion Protein

Choose a NanoLuc® Fusion Vector. We offer a range of vectors designed for simple generation of NanoLuc® luciferase N- or C-terminal fusion proteins with CMV-driven expression.

View NanoLuc® Fusion Vectors

We also offer vector systems containing NanoLuc® luciferase gene fused to the C-terminus of the HIF1A or NRF2 proteins. These vectors are designed and tested for stability studies involving these two regulatory proteins.

View NanoLuc® Stability Sensors

Step 2: Choose a Detection Reagent

For standard assays, the Nano-Glo® Luciferase Assay provides a simple, "add-and-read method for measuring NanoLuc® fusion protein abundance.

View Nano-Glo® Luciferase Assay System

Long-term live cell detection of NanoLuc® luciferase is possible with new Nano-Glo® Live Cell Reporter Substrates under development. Please inquire using the link below if you are interested in testing these substrates.

Please Inquire
GloMax Discover Instrument

The Nano-Glo® 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
Biosensors

BRET-based biosensors to detect signaling events inside and outside of the cell.

Biosensors
Gene Regulation & Cell Signaling

Greater sensitivity and versatility for traditional gene regulation assays.

Gene Regulation & Cell Signaling
Imaging

Sensitive, bright bioluminescent imaging in cells and whole animals.

Imaging
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