A novel human serum protein with a molecular mass of 87,000 daltons was purified to homogeneity and subjected to amino acid sequence analyses. These sequences were used to design oligonucleotide primers and to isolate a full-length cDNA. The amino acid sequence encoded by the cDNA shares strong similarity to albumin family members and shares the characteristic pattern of Cys residues observed in this family. In addition, the gene maps to chromosome 4 as do other members of the albumin gene family. Based upon these observations, we conclude that the 87,000-dalton protein, which we designate afamin (AFM), is the fourth member of the albumin family of proteins. Afamin cDNA was stably transfected into Chinese hamster ovary cells and recombinant protein (rAFM) was purified from conditioned medium. Both rAFM and AFM purified from human serum react with a polyclonal antibody that was raised against a synthetic peptide derived from the deduced amino acid sequence of AFM.