To aid in evaluating the functional significance of various domains of the hepatocyte plasma membrane, we have developed monoclonal antibodies that react with three different antigens on the surface of rat hepatocytes. One antigen is present on the sinusoidal-lateral plasma membrane but is absent from the bile canalicular membrane. On the basis of apparent molecular weight and ability to bind a desialylated glycoprotein, this antigen may be the asialoglycoprotein receptor. It was also identified immunocytochemically on the sinusoidal-lateral plasma membrane of human hepatocytes. The second antigen is present only on the bile canalicular membrane, whereas the third is present on the entire cell surface of rat hepatocytes. Our monoclonal antibodies may be useful in investigations of the sorting mechanisms of hepatocyte membrane proteins and the pathogenesis of certain liver diseases.