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Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells
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Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells

Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells

DOI:
10.3791/54899-v

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09:34 min

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December 10, 2016

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1Lund University Diabetes Centre,Lund University, 2INSERM UMR-1060, CarMeN Laboratory, Lyon 1 University, INRA U1235, INSA of Lyon,Rockefeller and Charles Merieux Lyon-Sud Medical Universities

Chapters

  • 00:05Title
  • 01:42Fixation and Blocking of Cells
  • 03:46Primary Antibodies and Proximity Ligation Assay Probes
  • 06:09Amplification and Imaging Preparation
  • 07:48Results: In Situ Proximity Ligation Assay in HuH7 Cells
  • 08:52Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a procedure to visualize and quantify with high sensitivity the endogenous interactions between the endoplasmic reticulum and mitochondria in fixed cells. The protocol features an optimized in situ proximity ligation assay targeting the inositol 1,4,5-triphosphate receptor/glucose-regulated protein 75/voltage-dependent anion channel/cyclophilin D complex at the mitochondria-associated membrane interface.

Tags

Keywords: Endoplasmic ReticulumMitochondriaIn Situ Proximity Ligation AssayFixed CellsVDAC1IP3R1Mitochondrial Associated MembraneImmunofluorescenceBiomedical ResearchCell FixationCell PermeabilizationBlockingImmunostaining

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