Preparation of a Total Membrane Fraction from 3T3-L1 Adipocytes.

Authors :: Sadler JB
Lamb CA
Gould GW
Bryant NJ
Source :: Cold Spring Harbor protocols [Cold Spring Harb Protoc] 2016 Mar 01; Vol. 2016 (3), pp. pdb.prot083675. Date of Electronic Publication: 2016 Mar 01.
Publication Year :: 2016
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.<br /> (© 2016 Cold Spring Harbor Laboratory Press.)