Quantitation of aspartate aminotransferase isoenzymes after electrophoretic separation.
A scheme for the quantitative detection of aspartate aminotransferase isoenzymes and multiple forms after electrophoretic separation is described. Glutamate generated from the aminotransferase reaction is quantitated by using the glutamate dehydrogenase/diaphorase-coupled enzyme system to form a formazan dye. Product inhibition of aspartate aminotransferase by oxaloacetate is prevented by including oxaloacetate decarboxylase in the overlay reagent. Results compare favorably with those of an immunochemical precipitation procedure. The method can also be used to detect quantitatively subforms and atypical forms (genetic variants, immunoglobulin-enzyme complexes) of aspartate aminotransferase.
D000818:Animals; D001219:Aspartate Aminotransferases / Q000032:analysis*; D002262:Carboxy-Lyases; D011232:Chemical Precipitation; D008058:Dihydrolipoamide Dehydrogenase; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology; D005969:Glutamate Dehydrogenase; D006801:Humans; D007120:Immunochemistry; D007527:Isoenzymes / Q000032:analysis*; D008928:Mitochondria / Q000201:enzymology; D051381:Rats
D007527:Isoenzymes; D005969:Glutamate Dehydrogenase; D008058:Dihydrolipoamide Dehydrogenase; D001219:Aspartate Aminotransferases; D002262:Carboxy-Lyases; C021916:oxaloacetate decarboxylase