Description
Overview of Nano-Glo® Live Cell Detection
Live-cell, nonlytic assays using the Nano-Glo® Live Cell Assay System are limited to ≤2 hours because of signal decay. The Nano-Glo® Endurazine™ and Vivazine® Live Cell Substrates provide alternative live-cell-detection methods for NanoLuc® and NanoBiT® luciferases that enable nonlytic assays for periods lasting several hours to days. For both substrates, a slow rate of ester hydrolysis leads to the steady release of furimazine throughout the experiment, a process catalyzed by cellular esterases. Once formed, furimazine serves as a substrate for NanoLuc® and NanoBiT® luciferases.
The Vivazine® substrate typically shows increased brightness but also an increased rate of signal decay compared to the Endurazine™ substrate. The Endurazine™ substrate will provide the maximum signal stability but lower initial signal intensity compared to the other Nano-Glo® Live Cell Substrates. Evaluate both substrates to determine which combination of signal intensity and stability is most suitable for your experiment [e.g., Nano-Glo® Extended Live Cell Substrate Trial Pack (Cat.# N2590)].
Choose the Best Combination of Signal Intensity and Stability
A plasmid encoding NanoLuc® luciferase expressed from the PGK promoter was transiently transfected into HEK293 cells and luminescence signal intensity was measured as indicated.
Assay Reporter Activity for Days
A plasmid encoding NanoLuc® luciferase expressed from the HSV-TK promoter was transiently transfected into HEK293 cells and luminescent signal intensity measured for 72 hours.
