Unit

UNIT 4.3 Subcellular Fractionation of Tissue Culture Cells

  1. Fernando Aniento1,
  2. Jean Gruenberg2

Published Online: 1 AUG 2004

DOI: 10.1002/0471140864.ps0403s32

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Aniento, F. and Gruenberg, J. 2004. Subcellular Fractionation of Tissue Culture Cells. Current Protocols in Protein Science. 32:4.3:4.3.1–4.3.21.

Author Information

  1. 1

    University of Valencia, Valencia, Spain

  2. 2

    University of Geneva Sciences II, Geneva, Switzerland

Publication History

  1. Published Online: 1 AUG 2004
  2. Published Print: MAY 2003

Abstract

Cell fractionation techniques include some of the most important and widely used analytical tools in cell and molecular biology, and are essential for the development of cell-free assays that reconstitute complicated cellular processes. In addition to simple gradient systems, this unit discusses the immuno-purification of organelles, in particular endosomes. As antigens, purification can be achieved using endogenous or ectopically expressed proteins, provided that appropriate antibodies are available. Alternatively, tagged proteins can be used, when combined with anti-tag antibodies. Now that sequencing of the genomes of several organisms has been completed, biochemical strategies, and in particular fractionation and in vitro transport assays, are more necessary than ever to study the numerous protein and protein complexes that are being discovered.