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生物化学
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Quantitative Immunofluorescence to Measure Global Localized Translation
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生物化学
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JoVE 杂志 生物化学
Quantitative Immunofluorescence to Measure Global Localized Translation

Quantitative Immunofluorescence to Measure Global Localized Translation

DOI:
10.3791/55909-v

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09:13 min

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August 22, 2017

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1Centre de Recherche sur le Cancer de l’Université Laval, Faculté de Médecine, Département de Biologie moléculaire, biochimie médicale et pathologie,Université Laval, 2CRCHU de Québec: L’Hôtel-Dieu de Québec

Chapters

  • 00:05Title
  • 00:45Cell Immunostaining
  • 03:34Image Acquisition
  • 04:49Signal Quantification
  • 06:50Results: Proper Puromycin Treatment and Analysis in MRC-5 and Other Cell Types
  • 08:16Conclusion

Summary

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This manuscript describes a method to visualize and quantify localized translation events in subcellular compartments. The approach proposed in this manuscript requires a basic confocal imaging system and reagents and is rapid and cost-effective.

Tags

Keywords: Quantitative ImmunofluorescenceLocalized TranslationCell AdhesionPuromycinAnti-puromycin AntibodyFluorophore-conjugated Secondary AntibodiesCell MigrationCell AdhesionEarly Drug ResponseMRC-5 CellsCycloheximideFormaldehyde FixationTriton X-100 PermeabilizationBSA BlockingTween-20 Washing

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