Esterase 1 (Es-1) is a sexually dimorphic 65-kDa glycoprotein present in plasma and other murine tissues able to hydrolyze a variety of esters including fatty acid esters of estradiol. Like most other carboxylesterases, its function is unknown. To gain insight into the function of Es-1 and by analogy other carboxylesterases, we have examined the developmental regulation of Es-1 in the mouse and have looked for the presence of related proteins in the plasma of other species. Northern blot analysis of total RNA from the livers of mice of various ages using a 32P-labeled 470-bp Es-1 cDNA probe showed clear postpartum induction with no detectable Es-1 mRNA in fetal liver. Similarly, immunoblotting after sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an affinity-purified rabbit antibody to Es-1 showed no cross-reacting proteins in the plasma until after birth. Northern blot analysis of total RNA from a variety of adult mouse tissues showed the presence of substantial levels of Es-1 mRNA only in liver with lower levels in kidney, testes, and ovaries. Liver mRNA and plasma protein levels rose in parallel attaining full adult levels between 15 and 20 days of age. When plasma proteins were electrophoresed on 7% polyacrylamide gels under nondenaturing conditions, the antibody to Es-1 recognized a low mobility protein in mouse, rat, human, baboon, guinea pig, bovine, horse, and canine but not in chicken plasma. Consistent with the immunoblotting results, the Es-1 cDNA probe hybridized to restriction fragments from human, monkey, rat, and rabbit as well as mouse genomic DNA but not from chicken DNA indicating conservation of the esterase (or esterase-like) gene in mammalian species. The low mobility antigens in mouse and human plasma appeared also to cross-react with antibodies to human thyroglobulin, although antibodies to human thyroglobulin did not appear to recognize Es-1 under these conditions.