Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | Unit 5.4 |
| Journal | Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.] |
| Volume | Chapter 5 |
| State | Published - May 2001 |
ASJC Scopus subject areas
- Cell Biology