Description
Use Either Firefly or NanoLuc® Luciferase as the Primary Reporter or use Both as Dynamic Reporters
Use the Nano-Glo® Dual-Luciferase® Reporter (NanoDLR™) Assay in different configurations to suit your assay design.
NanoLuc® Luciferase as Primary Reporter
The brightest, most responsive format.
Best for: Low cell numbers, plate scale-up or challenging cell lines.
Reporter Constructs: NlucP Experimental Reporter; Fluc Control.
Highlighted data shows luminescence values for HEK293 cells transfected with NFkBNanoLuc® PEST vector and TK-Fluc control at 10:1 ratio. Cells were treated with TNFα and reporter activity measured using the NanoDLR™ assay 4 hours post-treatment.
Firefly Luciferase as Primary Reporter
Use existing firefly luciferase constructs, and add NanoLuc® luciferase for a more robust control.
Best for: Standard reporter assay formats. The NanoLuc® luciferase control reporter is up to 1000X brighter than Renilla luciferase, so you can use less control DNA.
Reporter Constructs: FlucP Experimental Reporter; Nluc Control
Highlighted data shows luminescence values for HEK293 cells transfected with a NFkB response element Fluc-PEST vector and a TK-Nanoluc® control vector at 10:1 ratio. Cells were treated with TNFα and reporter activity measured using the NanoDLR™ assay 4 hours post-treatment.
Two Primary Reporters
Measure two different biological responses simultaneously.
Best for: Measuring 2 pathways at once for maximum data; multiplexing protein stability and genetic reporter assays.
Reporter Constructs: NlucP Experimental Reporter; FlucP Experimental Reporter; Nluc protein stability reporter.
In this example, the NanoDLR™ Assay was used to measure protein dynamics and gene activation in the same sample. HEK293 cells were transiently transfected with the pNLF1-HIF1A[CMV/neo] fusion construct, diluted 1:1000 into the Hypoxia Response Element Vector, pGL4.42[luc2P/HRE/Hygro]. After 18 hours, cells were stimulated with varying doses of 1,10-phenanthroline and assayed for expression of the firefly luciferase transcriptional reporter and the HIF1a-NanoLuc® fusion protein using the Nano-Glo® Dual-Luciferase® Reporter (NanoDLR™) Assay at the indicated time points.
Coincidence Reporter System
Express both reporters from a single open reading frame.
Best for: Distinguishing false hits in HTS from compounds truly affecting expression of target gene.
Reporter Construct: Firefly-P2A-NanoLuc®-PEST construct (pNLCoI Vectors).
