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Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
JoVE Journal
Biology
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JoVE Journal Biology
Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
DOI:

09:59 min

March 16, 2017

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Chapters

  • 00:05Title
  • 00:39Live Cell Imaging by TIRFM
  • 02:48Imaging SEP to Determine Subcellular Localization
  • 04:39Imaging Single Vesicle Insertion Events in Live Cells
  • 05:44Image Analysis and Data Processing
  • 08:17Results: Monitoring the Localization, Expression, and Trafficking of Membrane Proteins
  • 09:30Conclusion

Summary

Automatic Translation

Labeling the extracellular domain of a membrane protein with a pH sensitive fluorophore, superecliptic pHluorin (SEP), allows subcellular localization, expression, and trafficking to be determined. Imaging SEP-labeled proteins with total internal reflection fluorescence microscopy (TIRFM) enables the quantification of protein levels in the peripheral ER and plasma membrane.

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