Journal
/
Biochemistry
/
Quantitative Immunofluorescence to Measure Global Localized Translation
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Quantitative Immunofluorescence to Measure Global Localized Translation

Quantitative Immunofluorescence to Measure Global Localized Translation

DOI:
10.3791/55909-v

•

09:13 min

•

August 22, 2017

•

,
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

Automatic Translation

Automatic Translation

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

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Genome-wide Quantification of Translation in Budding Yeast by Ribosome Profiling

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions

Ubiquitin Chain Analysis by Parallel Reaction Monitoring

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA

A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion

Förster Resonance Energy Transfer Mapping: A New Methodology to Elucidate Global Structural Features

Read Article