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pNLF1-HIF1A [CMV/neo] Vector

by Promega
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  • The rate of protein turnover is tightly regulated for many signaling proteins involved in oncogenesis and response to cellular stress. Protein stabilization and subsequent accumulation occurs in response to changing cellular conditions resulting in activation of downstream transcriptional events. The NanoLuc® Stability Sensors are ready-to-use vector systems that utilize the advantages of the NanoLuc® luciferase reporter to enable stability studies of two key signaling proteins, HIF1A and NRF2, providing a method to directly measure this primary signaling event.

    HIF1A Vector System: The HIF1A Vector System enables simple quantification of intracellular HIF1A protein levels to study the dynamics of this signaling protein in mediating cellular response to hypoxia. It contains a vector encoding NanoLuc® fused to the C-terminus of the HIF1A protein under control of the CMV promoter plus Transfection Carrier DNA for titrating intracellular fusion protein expression.

    NRF2 Vector System: The NRF2 Vector System enables simple quantification of intracellular NRF2 protein levels to study the dynamics of this signaling protein in mediating cellular response to oxidative stress. It contains a vector encoding NanoLuc® fused to the C-terminus of the NRF2 protein under the control of the CMV promoter, a pKEAP1-expressing vector for proper regulation of intracellular NRF2 levels and Transfection Carrier DNA for titratable intracellular fusion protein expression.

    Applications

    • Compound screening.
    • Hypoxia signaling.
    • Oxidative stress signaling.

    Sequence Information

    pNLF1-HIF1A [CMV/neo] Vector GenBank® Accession Number KF853601 and vector sequence text file.

    pNLF1-NRF2 [CMV/neo] Vector GenBank® Accession Number KF853602 and vector sequence text file
    (pKEAP1 [CMV/Hygro] Vector GenBank® Accession Number KF853604 and vector sequence text file).

    Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.
    Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.
    Rapid stabilization and accumulation of NRF2 monitored with NRF2-NanoLuc® fusion protein.
    Rapid stabilization and accumulation of NRF2 monitored with NRF2-NanoLuc® fusion protein.
  • Protocols

    Complete Protocol

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    pNLF1-HIF1a [CMV/neo] Vector

    PDF (205 KB)

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    pNLF1-NRF2 [CMV/neo] Vector

    PDF (203 KB)

  • Certificate of Analysis

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    Storage Conditions

    -30C TO -10C

    Use Restrictions

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

  • Resources

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