pFC32A Nluc CMV-Hygro Flexi Vector
Generate N-terminal fusions with NanoLuc® luciferase using the pFN31A and pFN31K Nluc CMV-neo Flexi® Vectors while the pFC32A and pFC32K Nluc CMV-neo Flexi® Vectors create C-terminal Nluc fusion proteins. These vectors are part of the Flexi® Vector Cloning System that uses a directional cloning method based on two rare-cutting restriction enzymes, SgfI and PmeI, that provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi® Vectors without the need to resequence. All vectors contain a mammalian selectable marker to create a stable line.
The small size (19.1kDa) and extreme brightness of NanoLuc® luciferase (Nluc; about 100-fold brighter than either firefly [Photinus pyralis] or Renilla reniformis) make it an ideal protein fusion partner. When visualizing intracellular protein dynamics, the bright NanoLuc® reporter reduces imaging exposure times without the need for repeated sample excitation, which can result in cytotoxic artifacts.
NanoLuc® fusion proteins can be used in a variety of applications including: reporters of protein stability, probes for bioluminescent cell imaging (BLI) or as the donor signal in bioluminescent resonance energy transfer (BRET) applications for protein:protein or protein:small-molecule interaction studies. The brighter signal and blue-shifted emission spectrum from NanoLuc® luciferase result in less spectral overlap with fluorescent acceptors, resulting in better signal:background and dynamic range for BRET applications.
Additional NanoLuc® protein fusion vectors include the pNLF Vector series. Generate N- or C-terminal fusions to the full-length Nluc protein or attach secreted Nluc to the N-terminus of the protein of interest using traditional cloning with a multiple cloning site (MCS).
- Compound screening.
- Protein:protein interactions.
- Protein:small-molecule interactions.
- Bioluminescent imaging.
- Protein stability.
- Hall, M.P. (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chemical Biology 7(11), 1848–57.
Certificate of AnalysisLookup Certificate of Analysis
-30C TO -10C
For Research Use Only. Not for Use in Diagnostic Procedures.