Skip to content

pFC32K Nluc CMV-neo Flexi Vector

by Promega
Login to view price.
Size
  • Overview
  • Protocols
  • Specifications
  • Resources
  • 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).

    Applications

    • Compound screening.
    • Protein:protein interactions.
    • Protein:small-molecule interactions.
    • Bioluminescent imaging.
    • Protein stability.

    Reference

    1. Hall, M.P. (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chemical Biology 7(11), 1848–57.

    Sequence Information

    pFN31A Nluc CMV-Hygro Flexi® Vector GenBank® Accession Number KF793052 and vector sequence text file.

    pFN31K Nluc CMV-neo Flexi® Vector GenBank® Accession Number KF793053 and vector sequence text file.

    pFC32A Nluc CMV-Hygro Flexi® Vector GenBank® Accession Number KF811455 and vector sequence text file.

    pFC32K Nluc CMV-neo Flexi® Vector GenBank® Accession Number KF811456  and vector sequence text file.

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources

    Articles

    Citations