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pGL4.42[luc2P/HRE/Hygro] Vector

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • Creating a cell line with an indicator of a functional signaling pathway is useful for deciphering the components in a signaling pathway. These tools are made by insertion of multiple repeats of a response element upstream of a minimal promoter (minP). Promega has designed vectors that report the activity of a variety of pathways using the optimized luc2 firefly luciferase gene in the pGL4 backbone. These vectors also have a hygromycin resistance selectable marker, allowing use either in transient transfection experiments or for selection of a stable cell line. In addition, Some of these pre-designed response element vectors also are available stable cell lines (GloResponse™ Cell Lines) to speed your research.

    There are more vectors for measuring cytokine signaling pathways and additional signaling pathways.

    Applications

    • Cell transcription.
    • Virus-cell interactions.
    • Compound screening.
    • Post-translational modifications.
    • GPCR signaling.
    • Cell signaling.

    About the pGL4 Reporter Vectors

    The pGL4 Vectors offer increased reporter gene expression with codon optimization of synthetic genes for mammalian expression and reduced background and risk of expression artifacts with removal of cryptic DNA regulatory elements and transcription factor binding sites. Firefly luciferase has options that offer improved temporal response with the destabilized Rapid Response™ luciferase genes.

    Sequence Information

    pGL4.37[luc2P/ARE/Hygro] Vector GenBank® Accession Number JQ858521 and vector sequence text file.

    pGL4.38[luc2P/p53 RE/Hygro] Vector GenBank® Accession Number JQ858522 and vector sequence text file.

    pGL4.39[luc2P/ATF6 RE/Hygro] Vector GenBank® Accession Number JQ858519 and vector sequence text file.

    pGL4.40[luc2P/MRE/Hygro] Vector GenBank® Accession Number JQ858515 and vector sequence text file.

    pGL4.41[luc2P/HSE/Hygro] Vector GenBank® Accession Number JQ858520 and vector sequence text file.

    pGL4.42[luc2P/HRE/Hygro] Vector GenBank® Accession Number JQ858518  and vector sequence text file.

    pGL4.43[luc2P/XRE/Hygro] Vector GenBank® Accession Number JQ858513 and vector sequence text file.

    Activator/Pathway Transcription Factor Binding Site Vector Name Example Data
    Oxidative stress Nrf2 Antioxidant Response Element (ARE) pGL4.37[luc2P/ARE/Hygro]
    Representative data for pGL4.37[luc2P/ARE/Hygro] Vector in HEK293 cells upon stimulation with tBHQ or D,L-Sulforaphane. HEK293 cells were transiently transfected with pGL4.37[luc2P/ARE/Hygro] and pGL4.75[hRluc/CMV] Vectors and assayed in 96-well format after 18 hours stimulation with tBHQ or D,L-Sulforaphane as described. Firefly luciferase luminescence normalized to the Renilla luciferase control is shown, with error bars indicating the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagents, using a GloMax® 96 instrument with a 0.5 second integration time.
    DNA damage

    p53

    p53 response element (p53 RE) pGL4.38[luc2P/p53 RE/Hygro]
    Representative data for pGL4.38[luc2P/p53 RE/Hygro] Vector in U2OS cells upon stimulation with doxorubicin, etoposide, nutlin-3 and mitomycin c. U2OS cells were transiently transfected with the pGL4.38[luc2P/p53 RE/Hygro] and pGL4.75[hRluc/CMV] Vectors, and assayed in 96-well format after 18 hours stimulation with doxorubicin, nutlin-3, and etoposide, or after 40 hours with mitomycin c as indicated in the protocol. Firefly luciferase luminescence normalized to the Renilla luciferase control is shown, with error bars indicating the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagents, using a GloMax® 96 instrument with a 0.5 second integration time.
    Endoplasmic reticulum stress ATF6 Activating Transcription Factor 6 response element (ATF6 ERSE) pGL4.39[luc2P/ATF6 RE/Hygro]
    Representative data for pGL4.39[luc2P/ATF6 RE/Hygro] Vector in HeLa cells upon stimulation with tunicamycin. HeLa cells were transiently transfected with pGL4.39[luc2P/ATF6 RE/Hygro] and pGL4.75[hRluc/CMV] Vectors and assayed in 96-well format after 18 hours stimulation with tunicamycin as indicated in the protocol. Firefly luciferase luminescence normalized to the Renilla luciferase control is shown, with error bars indicating the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagents, using a GloMax® 96 instrument with a 0.5 second integration time.
    Heavy metal stress MTF1 Metal Regulatory Element (MRE) pGL4.40[luc2P/MRE/Hygro]
    Representative data for pGL4.40[luc2P/MRE/Hygro] Vector in HepG2 cells upon stimulation with ZnSO4. HepG2 cells were transiently transfected with pGL4.40[luc2P/MRE/Hygro] and pGL4.75[hRluc/CMV] Vectors and assayed in 96-well format after six hours stimulation with ZnSO4 as indicated in the protocol. Firefly luciferase luminescence normalized to the Renilla luciferase control is shown. Error bars indicate the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagents, using a GloMax® 96 instrument with a 0.5 second integration time.
    Heat shock HSF1 Heat Shock Element (HSE) pGL4.41[luc2P/HSE/Hygro]
    Representative data for pGL4.41[luc2P/HSE/Hygro] Vector in HepG2 cells upon stimulation with 17-AAG or CdCl2. HepG2 cells were transiently transfected with pGL4.41[luc2P/HSE/Hygro] and pGL4.75[hRluc/CMV] Vectors and assayed in 96-well format after six hours stimulation with 17-AAG or CdCl2 as indicated. Firefly luciferase luminescence normalized to the Renilla luciferase control is shown. Error bars indicate the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagents, using a GloMax® 96 instrument with a 0.5 second integration time.
    Hypoxia Hif1? Hypoxia Response Element (HRE) pGL4.42[luc2P/HRE/Hygro]
    Representative data for pGL4.42[luc2P/HRE/Hygro] Vector in HEK293 cells upon stimulation with 1,10-phenanthroline. HEK293 cells were transiently transfected with pGL4.42[luc2P/HRE/Hygro] and pGL4.75[hRluc/CMV] Vectors, and assayed in 96-well format after five hours stimulation with 1,10-phenanthroline as indicated in the protocol. Firefly luciferase luminescence is shown, normalized to untreated cells, with error bars indicating the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagent, using a GloMax® 96 instrument with a 0.5 second integration time.
    Xenobiotic stress

    AhR

    Xenobiotic Responsive Element (XRE) pGL4.43[luc2P/XRE/Hygro]
    Representative data for pGL4.43[luc2P/XRE/Hygro] Vector in HepG2 cells upon stimulation with TCDD. HepG2 cells were transiently transfected with pGL4.43[luc2P/XRE/Hygro] and pGL4.75[hRluc/CMV] Vectors and assayed in 96-well format after 24 hours stimulation with TCDD as indicated in the protocol. Firefly luciferase luminescence normalized to the Renilla luciferase control is shown, with error bars indicating the S.E.M. for six replicates. Luminescence was detected after addition of Dual-Glo® reagents, using a GloMax® 96 instrument with a 0.5 second integration time.
  • Protocols

    Complete Protocol

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    pGL4.37[luc2P/ARE/Hygro] Vector Protocol

    PDF (198 KB)

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    pGL4.38[luc2P/p53 RE/Hygro] Vector Protocol

    PDF (206 KB)

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    pGL4.39[luc2P/ATF6 RE/Hygro] Vector Protocol

    PDF (192 KB)

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    pGL4.40[luc2P/MRE/Hygro] Vector Protocol

    PDF (212 KB)

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    pGL4.41[luc2P/HSE/Hygro] Vector Protocol

    PDF (197 KB)

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    pGL4.42[luc2P/HRE/Hygro] Vector Protocol

    PDF (194 KB)

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    pGL4.43[luc2P/XRE/Hygro] Vector Protocol

    PDF (215 KB)

  • 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