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Detecting Protein Subcellular Localization by Green Fluorescence Protein Tagging and 4′,6-Diamidino-2-phenylindole Staining in Caenorhabditis elegans
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Biologia
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JoVE Journal Biologia
Detecting Protein Subcellular Localization by Green Fluorescence Protein Tagging and 4′,6-Diamidino-2-phenylindole Staining in Caenorhabditis elegans

Detecting Protein Subcellular Localization by Green Fluorescence Protein Tagging and 4′,6-Diamidino-2-phenylindole Staining in Caenorhabditis elegans

DOI:
10.3791/57914-v

•

09:36 min

•

July 30, 2018

•

, ,
1Department of Science,Borough of Manhattan Community College/CUNY

Capítulos

  • 00:04Título
  • 00:26Heat Stress
  • 02:46Ethanol Fixation and DAPI Staining
  • 04:10Acetone Fixation
  • 05:21Microscopic Imaging
  • 07:56Results: Visualization of C. elegans Exl-1 Subcellular Localization through GFP Tagging and DAPI Staining
  • 09:07Conclusion

Summary

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Tadução automática

This protocol demonstrates how to track a protein nuclear translocation under heat stress by using a green fluorescence protein (GFP) fusion protein as a marker and 4',6-diamidino-2-phenylindole (DAPI) staining. The DAPI staining protocol is fast and preserves the GFP and protein subcellular localization signals.

Tags

Keywords: Protein Subcellular LocalizationGFP TaggingDAPI StainingCaenorhabditis ElegansCell BiologyTranslational ConstructExtrachromosomal ArrayGamma RadiationTransgenic LineStress AssayHeat StressM9 BufferSynchronizationL4 Larvae

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