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Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence
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JoVE Journal Cancer Research
Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence

Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence

DOI:
10.3791/64920-v

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06:25 min

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February 10, 2023

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, ,
1Division of Oncology, Department of Medicine, Siteman Cancer Center,Washington University School of Medicine

Chapters

  • 00:05Introduction
  • 01:00Plating, Pulsing, Fixation, Permeabilization, and Blocking of Cells
  • 03:59Immunostaining with the IdU Antibody
  • 05:15Results: IdU-Induced Replication Stress
  • 05:41Conclusion

Summary

Automatic Translation

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Here, we describe an immunofluorescence-based method to quantify the levels of single-stranded DNA in cells. This efficient and reproducible method can be utilized to examine replication stress, a common feature in several ovarian cancers. Additionally, this assay is compatible with an automated analysis pipeline, which further increases its efficiency.

Tags

Keywords: Replication StressSingle-stranded DNAImmunofluorescenceOvarian CancerBRCA1BRCA2Cell LinesBiomarkerDNA RepairHigh-throughputQuantificationProtocol

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