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Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells
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Biochemistry
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JoVE Journal Biochemistry
Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells

Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells

DOI:
10.3791/55747-v

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12:03 min

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July 23, 2017

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, ,
1Queensland Alliance for Agriculture & Food Innovation,The University of Queensland, 2Centre for Comparative Genomics,Murdoch University

Chapters

  • 00:00Title
  • 00:28Dissection of the Gut Epithelium from R. microplus
  • 01:56Epithelial Cell Dissociation
  • 03:36Isolation of Single Epithelial Cells Using a Percoll Gradient
  • 06:47Cell Surface Biotinylation and the Isolation of Biotinylated Surface Proteins
  • 09:44Representative Results
  • 11:17Conclusion

Summary

Automatic Translation

Automatic Translation

A modified density centrifugation gradient-based methodology was utilized to isolate epithelial cells from Rhipicephalus microplus gut tissue. Surface-bound proteins were biotinylated and purified through streptavidin magnetic beads for utilization in downstream applications.

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Keywords: Biotinylated Cell Surface ProteinsRhipicephalus MicroplusEpithelial Gut CellsPercoll GradientCell DisassociationCell PurificationStreptavidin Magnetic Beads

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