Unit

UNIT 3.42 Isolation of Primary Cilia by Shear Force

  1. Kimberly A. P. Mitchell

Published Online: 1 JUN 2013

DOI: 10.1002/0471143030.cb0342s59

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Mitchell, K. A. P. 2013. Isolation of Primary Cilia by Shear Force. Current Protocols in Cell Biology. 59:3.42:3.42.1–3.42.9.

Author Information

  1. Department of Biology, Liberty University, Lynchburg, Virginia

Publication History

  1. Published Online: 1 JUN 2013
  2. Published Print: JUN 2013

Abstract

The cell's primary cilium is both a mechanical and chemical sensor involved in many signaling pathways. In order to ascertain protein enrichment in the primary cilium or study sub-ciliary localization of various proteins, it is advantageous to remove the primary cilium from the cell body. The protocol described here gives detailed instructions on purifying primary cilia by separating them from the cell body using shear force. This simple technique avoids using harsh purification conditions that may affect signaling proteins in the cilium or cause the ciliary membrane to disintegrate. In addition, as the cell body remains mostly intact, contamination of the isolated cilia by proteins from the cell body is minimized. This protocol is ideally suited for isolating cilia from renal cell lines, as primary cilia in these cells grow to greater lengths than in other cell types (up to 50-µm long in Xenopus A6 toad kidney cells as opposed to 1 to 5 µm in NIH3T3 fibroblast cells). Curr. Protoc. Cell Biol. 59:3.42.1-3.42.9. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • primary cilia;
  • isolation;
  • purification;
  • shear force