Radioimmunoassay of aspartate aminotransferase isoenzymes in human serum.
We describe the development of a sensitive, specific radioimmunoassay for the cytoplasmic and mitochondrial isoenzymes of human aspartate aminotransferase (L-aspartate:2-oxoglutarate aminotransferase; EC 2.6.1.1). Isoenzymes from human heart tissue were purified to homogeneity and used to raise high-titer antisera in rabbits. We partly purified the antisera by selective column chromatography. The Bolton-Hunter reagent was used to radioiodinate the isoenzymes. The assay requires 100 microL of serum, includes a solid-phase second-antibody separation, and can be completed in less than 3 h. There was no cross reactivity between the two isoenzymes. As little as 5 micrograms (50 pmol) of each aspartate aminotransferase can be measured per liter of serum.
D000917:Antibody Formation; D000918:Antibody Specificity; D001219:Aspartate Aminotransferases / Q000097:blood* / Q000276:immunology; D001666:Binding Sites, Antibody; D003593:Cytoplasm / Q000201:enzymology; D006801:Humans; D007457:Iodine Radioisotopes; D007527:Isoenzymes / Q000097:blood; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D011863:Radioimmunoassay
D007457:Iodine Radioisotopes; D007527:Isoenzymes; D001219:Aspartate Aminotransferases