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Immunology and Infection
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Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells

Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells

DOI:
10.3791/59697-v

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00:06 min

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August 27, 2019

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,
1Norfolk Academy, 2Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute,Yale School of Medicine

Chapters

  • 00:04Title
  • 00:28Cell Fixation, Immunofluorescence, and Hybridization, and Viral RNA Visualization
  • 04:48Results: Representative FISH and IF Kaposi s Sarcoma-Associated Herpesvirus (KSHV) Oligonucleotide Analyses
  • 05:52Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a protocol utilizing fluorescence in situ hybridization (FISH) to visualize multiple herpesviral RNAs within lytically infected human cells, either in suspension or adherent. This protocol includes quantification of fluorescence producing a nucleocytoplasmic ratio and can be extended for simultaneous visualization of host and viral proteins with immunofluorescence (IF).

Tags

Keywords: Quantitative FISHImmunofluorescenceKSHVHost-virus InteractionsViral Gene ProductsRNAProteinBiochemical AssaysCell CultureFixationPermeabilizationBlockingPrimary AntibodySecondary AntibodyFluorescent LabelingHybridizationOligonucleotidesDenaturation

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