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Large-Scale Purification of Porcine or Bovine Photoreceptor Outer Segments for Phagocytosis Assays on Retinal Pigment Epithelial Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
Large-Scale Purification of Porcine or Bovine Photoreceptor Outer Segments for Phagocytosis Assays on Retinal Pigment Epithelial Cells

Large-Scale Purification of Porcine or Bovine Photoreceptor Outer Segments for Phagocytosis Assays on Retinal Pigment Epithelial Cells

DOI:
10.3791/52100-v

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10:02 min

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December 12, 2014

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, , , ,
1INSERM, U968, 2Sorbonne Universités, UPMC Paris 06, UMR_S 968,Institut de la Vision, 3CNRS, UMR_7210, 4Department of Biological Sciences, Center for Cancer, Genetic Diseases and Gene Regulation,Fordham University

Chapters

  • 00:05Title
  • 02:01Set-up, Solution and Gradient Preparation
  • 03:20Tissue Collection and Retina Homogenization
  • 04:54Photoreceptor Outer Segment Fragment Isolation
  • 05:58POS Aliquoting Without and With Labeling
  • 07:48Results: Large-scale Purification of Photoreceptor Outer Segments
  • 09:28Conclusion

Summary

Automatic Translation

Automatic Translation

This article describes the protocol for the purification of photoreceptor outer segment fragments (POS) via ultracentrifugation from porcine/bovine retinae using homogenization and sucrose gradient centrifugation. This protocol allows the preparation of large stocks of POS aliquots, labeled or unlabeled, that can then be stored at -80 °C.

Tags

Keywords: Photoreceptor Outer SegmentsRetinal Pigment Epithelial CellsPhagocytosis AssayPorcineBovineSucrose GradientPhotopigmentFITC LabelingFluorescence QuantificationOxidative StressAging

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