Rapid preparation of a plasma membrane fraction from adipocytes and muscle cells: application to detection of translocated glucose transporter 4 on the plasma membrane.

The aim of this study was to establish a rapid preparation of plasma membrane from adipocytes and muscle cells to detect translocated glucose transporter (GLUT) 4. A plasma membrane fraction was prepared by sequential centrifugation with buffer containing detergents, and its purity was estimated by detecting insulin receptor beta-subunit (IRbeta). After insulin stimulus, GLUT4 translocation was observed in 3T3-L1 adipocytes and L6 myotubes. It was found that IRbeta and GLUT4 levels on the plasma membrane decreased in adipose and muscle with intake of a 29% lard diet for 14 weeks. Hence, this method should be useful for rapid preparation of the plasma membrane fraction.