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Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay
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Biochemistry
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JoVE Journal Biochemistry
Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay

DOI:
10.3791/55703-v

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13:10 min

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June 09, 2017

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, , , , ,
1Department of Pathology,Academic Medical Center, University of Amsterdam, Lymphoma and Myeloma Center Amsterdam (LYMMCARE)

Chapters

  • 00:05Title
  • 01:02Treatment of Cells and Induction of DNA Damage
  • 03:21Cytocentrifugation, Fixation, and Permeabilization
  • 05:35Blocking and Primary Antibody Incubation
  • 07:19PLA Probes, Ligation, and Amplification
  • 09:04Mounting and Imaging
  • 10:34Results: The PLA Assay Detects Direct Protein-protein Interactions in Suspension Cell Cultures
  • 12:03Conclusion

Summary

Automatic Translation

Automatic Translation

Here, it is demonstrated how the in situ Proximity Ligation Assay (PLA) can be used to detect and visualize the direct protein-protein interactions between ATM and p53 in suspension cell cultures exposed to genotoxic stress.

Tags

Keywords: DNA DamageProtein ComplexesProximity Ligation AssaySuspension Cell CulturesDNA RepairATM InhibitorNCSBV-173 CellsCell Cycle ArrestMicroscopy Slides

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