Gradient por electrophoresis, with special reference to alkaline phosphatase isoenzymes

Abstract

Gradient pore polyacrylamide gel electrophoresis is a useful tool in the separation of proteins. Polyacrylamide gels of continuously decreasing pore size are cast, and electrophoresis of the protein mixture is carried out for an extended period. The individual proteins migrate until they are excluded by the gel. At the exclusion limit they are focused into sharp bands exhibiting excellent resolution. A simple apparatus has been designed to cast gradient gels in 'disc' electrophoresis tubes, putting the method within the scope of the clinical laboratory. The method has been used successfully in the separation of the isoenzymes of alkaline phosphatase, giving much greater resolution than previous methods. Complete separation of the liver and bone isoenzymes is obtained. The method is also convenient in the analysis of dilute protein solutions such as CSF or urine as a much larger volume of sample can be used without any loss in resolution. Gradient pore polyacrylamide gel electrophoresis is a useful tool in the separation of proteins. Polyacrylamide gels of continuously decreasing pore size are cast, and electrophoresis of the protein mixture is carried out for an extended period. The individual proteins migrate until they are excluded by the gel. At the exclusion limit they are focused into sharp bands exhibiting excellent resolution. A simple apparatus has been designed to cast gradient gels in 'disc' electrophoresis tubes, putting the method within the scope of the clinical laboratory. The method has been used successfully in the separation of the isoenzymes of alkaline phosphatase, giving much greater resolution than previous methods. Complete separation of the liver and bone isoenzymes is obtained. The method is also convenient in the analysis of dilute protein solutions such as CSF or urine as a much larger volume of sample can be used without any loss in resolution.