Preparation of a Total Membrane Fraction from 3T3-L1 Adipocytes
- 1University of Glasgow, Glasgow G12 8QQ, United Kingdom;
- 2Cancer Research UK, London Research Institute, London EC1V 4AD, United Kingdom;
- 3University of York, Heslington, York YO10 5DD, United Kingdom
Abstract
The dynamic nature of insulin-sensitive glucose transporter isoform 4 (GLUT4) storage vesicles (GSVs) makes their characterization challenging. Fractionation techniques can facilitate isolation of GSVs from insulin-sensitive cells. In this protocol, we describe preparation of a total membrane fraction from 3T3-L1 adipocytes. The resulting pellet contains all membranes and allows for easier identification of membrane proteins, including the insulin-sensitive pool of GLUT4. A method for concentration of the soluble fraction is also included.
Footnotes
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↵4 Correspondence: nia.bryant{at}york.ac.uk
- © 2016 Cold Spring Harbor Laboratory Press
