UNIT 5.4 Analysis of the Association of Proteins with Membranes
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Cell Biology
How to Cite
Okamoto, T., Schwab, R., Scherer, P. and Lisanti, M. 2001. Analysis of the Association of Proteins with Membranes. Current Protocols in Cell Biology. 5:5.4:5.4.1–5.4.17.
- Published Online: 1 MAY 2001
- Published Print: JAN 2000
Proteins associate with membranes in a number of ways. Is the membrane an integral membrane protein? A peripheral protein? Does it have a glycosylphosphoinositol (GPI) tail? Is it associated with caveolae-derived plasma membrane domains? This unit describes methods for determining how a protein of interest associates with the membrane including extraction with alkaline carbonate, urea, high salt, or Triton X-114. GPI linkage is probed using PI-PLC cleavage. There is a method for detergent solubilization of Triton-X 100 insoluble integral and GPI-linked proteins. Caveolae membranes can be isolated using Triton X-100 or a detergent-free method.