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检测和可视化DNA损伤诱导的蛋白质复合物悬浮细胞培养使用邻近连接测定
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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
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检测和可视化DNA损伤诱导的蛋白质复合物悬浮细胞培养使用邻近连接测定

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

在这里，证明了原位邻近连接测定（PLA）如何可以用于检测和可视化暴露于基因毒性应激的悬浮细胞培养物中ATM和p53之间的直接蛋白质 - 蛋白质相互作用。

Tags

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

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