The Isolation, Detection, and Quantification of Glucose Transporters

UNCP Author/Contributor (non-UNCP co-authors, if there are any, appear on document)
Grover Kevin Locklear (Creator)
The University of North Carolina at Pembroke (UNCP )
Web Site:

Abstract: The purpose of this research was to select a method of glucose transporter study, which is used to isolate, detect, and quantify glut-4, the isoform of glucose transporters found in muscle. Our long term goal is to find the vital link between glucose transporters and diabetes. To do this, an antibody against glut-4 was supplied to us by East Carolina School of Medicine. Glut-4 was then isolated from homogenized rat skeletal muscle. Samples of brain, kidney, liver (none of which contain glut-4 glucose transporters) were taken as negative controls, while samples of adipose and cardiac muscle (which contain glut-4 glucose transporters) were taken as positive controls. The proteins of the each tissue-type were immobilized on nylon paper via dot blotting, which showed non-specific binding. Since the dot blot was over reactive, we employed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which would give better separation of the proteins. The proteins were then transferred from the gel to a nitrocellulose membrane. The primary glut-4 antibody, from East Carolina, was added to the membrane, which contained the immobilized proteins. The secondary glut-4 antibody complex for binding. When the substrate for the alkaline phosphatase enzyme was added, a color change indicated where the glut-4 glucose transporter was present.

Additional Information

Language: English
Date: 1993
Glucose Transporter, Glut-4, Diabetes, Adipose, Cardiac Muscle, Nitrocellulose Membrane, Antibody,

Email this document to