Calbindin-D28K (Ca-D28K) is a calcium-binding protein. In the kidney, Ca-D28K is present in the distal nephron, but not in the proximal nephron. This site-specific distribution in the kidney indicates that Ca-D28K is a potential marker for the differentiation of the distal nephron. In this study, we have examined the expression of Ca-D28K in 25 sporadic cases of chicken nephroblastomas. All cases of nephroblastomas were composed of atypical tubular structures, blastemal cells, and fibrous stroma in varying degrees of differentiation. Immunohistochemically in all nephroblastoma specimens, Ca-D28K was expressed in the epithelial cells of the subsets of tubular structures, but not in the blastema or the stroma. These results suggested that the tubuli in the nephroblastomas are able to differentiate into the phenotype of distal nephrons. Furthermore, Ca-D28K might develop as a novel diagnostic marker for nephroblastomas because this molecule is reported to be completely negative in other renal tumors, including renal cell carcinoma, chromophobe carcinomas, and oncocytoma.