Immunoquantitation of Cytochrome c in Cardiac Perfusate

Abstract An indirect three step ELISA has been assessed in order to detect the possible release of cytochrome c, a mitochondrial protein, from isolated and perfused guinea-pig heart. The ELISA described in this study is sufficiently sensitive and accurate to measure extracellular cytochrome c. the respiratory chain (1). Cytochrome c is a very well known protein and its structure has been determined in a great number of species including man (2,3). Being the only water-soluble component of the respiratory chain and for this reason and in view of its small molecular mass, it may be lost from the organelle under pathological conditions such as a mitochondria1 damage induced by ischemiareperfusion (4,5). We have set up an immunological method to quantitatively determine cytochrome c in heart effluent.