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Immunology and Infection

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

Article DOI: 10.3791/52100-v • 10:02 min • December 12th, 2014
December 12th, 2014 •
1,2,3, 1,2,3, 1,2,3, 4, 1,2,3
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

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Summary
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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.

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Large-scale Purification Porcine Bovine Photoreceptor Outer Segments Phagocytosis Assays Retinal Pigment Epithelial Cells Analysis Vital Functions RPE Cells In Vitro POS Elimination Rodent RPE Cells Porcine RPE Cells Bovine RPE Cells Human RPE Cells Experiments Variability Stock Of POS Isolation Frozen Storage Photopigments Orange Color Dim Red Light Retinae Collection Buffer Eyecups Homogenized Retinal Cell Suspension Sucrose Gradient Centrifugation Discrete Band Orange Color Characteristic Collection Of POS Samples
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