title: Preparation of a Total Membrane Fraction from 3T3-L1 Adipocytes.
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.
authors.affiliation: University of Glasgow, Glasgow G12 8QQ, United Kingdom;
authors.forename: Jessica B A
authors.initials: JB
authors.lastname: Sadler
authors.affiliation: Cancer Research UK, London Research Institute, London EC1V 4AD, United Kingdom;
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Lamb
authors.affiliation: University of Glasgow, Glasgow G12 8QQ, United Kingdom;
authors.forename: Gwyn W
authors.initials: GW
authors.lastname: Gould
authors.affiliation: University of York, Heslington, York YO10 5DD, United Kingdom.
authors.forename: Nia J
authors.initials: NJ
authors.lastname: Bryant
mesh_terms: D017667:Adipocytes / Q000648:ultrastructure*; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002460:Cell Line; D002462:Cell Membrane; D051379:Mice