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In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay
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Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay

In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay

DOI:
10.3791/60982-v

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08:56 min

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May 05, 2020

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, ,
1Division of Biological Sciences,University of Montana

Capítulos

  • 00:04Introduction
  • 00:49Dissection/Gonad Extrusion
  • 02:46Fixation/Blocking
  • 04:14Primary Antibody Incubation and PLA Probe (Secondary Antibody) Incubation
  • 05:21Ligation
  • 06:09Amplification and Final Washes
  • 06:45Coverslip Mounting
  • 07:15Results: Germlines Following Co-immunostaining or PLA
  • 08:10Conclusion

Summary

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This protocol demonstrates use of the proximity ligation assay to probe for protein-protein interactions in situ in the C. elegans germline.

Tags

Proximity Ligation AssayProtein-protein InteractionsC. ElegansGermlineDissectionImmunofluorescenceMicroscopy

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