The distribution of the isoenzymes of aspartate aminotransferase (AST, E.C. 2.6.1.1.) in equine tissues has been studied to ascertain whether the organ of origin may be identified when the total AST activity of serum is raised. Most tissues contain 3 isoenzymes of cytoplasmic origin (cAST) with isoelectric points of 5.6, 5.7 and 5.9, and one isoenzyme of mitochondrial (mAST) origin with an isoelectric point of 9. Serum from horses with azoturia contained an additional cytoplasmic subform with an isoelectric point of 5.8. This form could not be generated by ageing, freezing and thawing or binding of the enzyme to gamma globulins or lipids. The ratio of cAST to mAST when separated by ion exchange chromatography varies widely between tissues, with no cAST detection in lung and no mAST detection in serum. Ageing of equine muscle homogenates caused the formation of 2 artefactual subforms with isoelectric points of 6.0 and 6.1. It is concluded that, although the ratio of mitochondrial to cytoplasmic AST varies between tissues, there is no tissue specificity of either cytoplasmic or mitochondrial isoenzymes and examination of serum would not indicate the source.