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Analysis of SCAP N-glycosylation and Trafficking in Human Cells
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Journal JoVE Biologie
Analysis of SCAP N-glycosylation and Trafficking in Human Cells

Analysis of SCAP N-glycosylation and Trafficking in Human Cells

DOI:
10.3791/54709-v

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11:27 min

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November 08, 2016

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, , , , , ,
1Department of Radiation Oncology,The Ohio State University Comprehensive Cancer Center and College of Medicine

Chapitres

  • 00:05Titre
  • 01:05Detection of Endogenous SCAP Protein in Human Cells
  • 04:11Analysis of SCAP N-glycosylation in Human Cells
  • 07:15Detection of SCAP Trafficking from ER to Golgi Using GFP-SCAP in Human Cells
  • 09:07Results: SCAP Trafficking from ER to Golgi
  • 10:43Conclusion

Summary

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Traduction automatique

We describe a modified method for membrane fraction isolation from human cells and sample preparation for the detection of SCAP N-glycosylation and total protein by using western blot. We further introduce a GFP-labeling method to monitor SCAP trafficking using confocal microscopy. This protocol can be used in regular biology laboratories.

Tags

SCAPN-glycosylationTraffickingHuman CellsSREBPLipid MetabolismPNGaseU87 CellsCell Membrane FractionsNuclear ExtractsSDS-PAGEWestern BlotBradford Protein Assay

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