Fetuin, an abundant protein in fetal bovine plasma, is the bovine homolog of human alpha 2HS glycoprotein (alpha 2HS). In spite of numerous studies, the biological functions of both proteins remain elusive. We now report the remarkable 45Ca-binding activity of fetuin in fetal bovine serum that has been blotted on a membrane after electrophoresis. The Ca-binding ability of the purified protein was analyzed by equilibrium dialysis, which revealed that bovine fetuin had multiple Ca-binding sites, one of which had a Kd of 0.95 x 10(-4) M. It was also shown that the Ca-binding activity of fetuin was greater than that of albumin in serum of the bovine fetus at the late gestational stage. Since fetal bovine serum contains not only a high concentration of fetuin but also a high concentration of Ca, it is possible that fetuin functions to maintain high levels of Ca in fetal serum via normalization of the concentration of Ca2+ ions.