Rhodopsin purification from dark-adapted bovine retina

Methods Mol Biol. 2015:1271:21-38. doi: 10.1007/978-1-4939-2330-4_2.

Abstract

Structural and biophysical studies of rhodopsin have long depended upon the ready availability of bovine retina from the meat-packing industry and the relative ease of obtaining homogenous preparations of rhodopsin in the quantities and purities necessary for such study. Herein we present a modular purification methodology employing a combination of several strategies, beginning with sucrose gradient isolation of rod outer segments (ROS) from bovine retina, detergent solubilization of ROS, selective extraction of rhodopsin starting from this detergent-solubilized ROS, and further purification via size-exclusion chromatography, resulting in a preparation of high-purity rhodopsin at high concentration suitable for crystallization or other biophysical study.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Chromatography, Gel
  • Light*
  • Retina / metabolism*
  • Rhodopsin / metabolism*
  • Rod Cell Outer Segment / metabolism

Substances

  • Rhodopsin