Unit

UNIT 3.6 Isolation of Lysosomes from Tissues and Cells by Differential and Density Gradient Centrifugation

  1. John M. Graham

Published Online: 1 MAY 2001

DOI: 10.1002/0471143030.cb0306s07

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Graham, J. M. 2001. Isolation of Lysosomes from Tissues and Cells by Differential and Density Gradient Centrifugation. Current Protocols in Cell Biology. 7:3.6:3.6.1–3.6.21.

Author Information

  1. Liverpool John Moores University, Liverpool, United Kingdom

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 2000

Abstract

This unit covers the use of Percoll, Nycodenz, and iodixanol for purification of lysosomes from a light mitochondrial pellet or postnuclear supernatant. The first protocol describes isolation of lysosomes from rat liver using a Percoll gradient; it includes some comments on using brain and kidney tissue as source material. Alternatively, this protocol can be modified to enhance the separation of lysosomes by organelle density perturbation. Another alternative uses a discontinuous gradient of Nycodenz to purify lysosomes from a rat liver light mitochondrial pellet. A continuous iodixanol gradient, which can be used to purify other organelles in the light mitochondrial fraction, is yet another alternative for purifying lysosomes. Lysosomes can also be isolated from some of the more commonly used cultured cells. The unit also includes assays for common lysosome marker enzymes.