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NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

DOI:
10.3791/60567-v

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10:57 min

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January 12, 2020

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1Department of Immunology and Microbiology,University of Colorado Anschutz Medical Campus

Chapters

  • 00:04Title
  • 01:13Cell Passaging and Seeding
  • 02:51Preparation of Bacteria
  • 06:23Infection of Cells
  • 08:00Luciferase Analysis
  • 09:18Results: Luminescence Assay and RT-qPCR Analysis
  • 10:10Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol to quickly and easily measure nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation in cell lines expressing NF-κB::luciferase reporter constructs, via measurements of luminescence in the cell lysate. Additionally, gene expression is determined via RT-qPCR isolated from cells infected with Salmonella Typhimurium.

Tags

Keywords: NF-κBLuciferaseGene ExpressionSalmonellaCell CultureTranscription FactorInflammatory ResponseCytokinesChemokinesHeLa CellsReporter PlasmidCell StimulationCell CountingCell Suspension48-Well PlateBacterial CultureLB Agar Plates

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