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Analysis of Endocytic Uptake and Retrograde Transport to the Trans-Golgi Network Using Functionalized Nanobodies in Cultured Cells
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Biochemistry
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JoVE Journal Biochemistry
Analysis of Endocytic Uptake and Retrograde Transport to the Trans-Golgi Network Using Functionalized Nanobodies in Cultured Cells

Analysis of Endocytic Uptake and Retrograde Transport to the Trans-Golgi Network Using Functionalized Nanobodies in Cultured Cells

DOI:
10.3791/59111-v

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11:05 min

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February 21, 2019

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,
1Biozentrum,University of Basel

Chapters

  • 00:04Title
  • 01:03Bacterial Liquid Culture and Induction of Functionalized Nanobody Expression
  • 02:31Purification of Functionalized Nanobodies
  • 04:14Validation of Functionalized Nanobody Expression
  • 04:41Uptake of Functionalized Nanobodies by Cultured Cells
  • 08:03Results: Analysis of Endocytic Uptake and Retrograde Transport to the Trans-Golgi Network
  • 10:09Conclusion

Summary

Automatic Translation

Automatic Translation

Retrograde transport of proteins from the cell surface to the Golgi is essential to maintain membrane homeostasis. Here, we describe a method to biochemically analyze cell surface-to-Golgi transport of recombinant proteins using functionalized nanobodies in HeLa cells.

Tags

Keywords: NanobodyEndocytic UptakeRetrograde TransportTrans-Golgi NetworkCell Surface ReceptorsFluorescence MicroscopyElectron MicroscopyTyrosine Sulfation

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