Quantitative methods for measuring the insulin-regulatable glucose transporter (Glut4).
This review article describes various quantitation methods for the insulin-regulatable glucose transporter (Glut4). Several methods including reconstituted glucose transport, cytochalasin B binding assays, immunocytochemistry, immunoblots, ELISA, and the more recently developed exofacial labels are discussed. Since Glut4 translocates from an intracellular compartment to the plasma membrane in response to the action of insulin, it is of particular interest to measure Glut4 changes in the membrane fractions. Hence, the measurement of Glut4 commonly involves the isolation of cell membranes using subcellular fractionation in combination with one of the quantitation methods. The limitations of each quantitation method due to the use of subcellular fractionation are discussed in this article. As well, the advantages and disadvantages in terms of isoform specificity and technical difficulties of each method are presented.
Li
W M
WM
Division of Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, Canada.
McNeill
J H
JH
D000345:Affinity Labels; D000818:Animals; D001692:Biological Transport; D003571:Cytochalasin B / Q000378:metabolism; D051275:Glucose Transporter Type 4; D006801:Humans; D007158:Immunologic Techniques; D009004:Monosaccharide Transport Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D009124:Muscle Proteins
D000345:Affinity Labels; D051275:Glucose Transporter Type 4; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; C495692:SLC2A4 protein, human; D003571:Cytochalasin B