A prospective nested case-control study of the relationship between prediagnostic serum levels of 1,25-dihydroxyvitamin D and subsequent development of malignant melanoma was done to test the hypothesis that increased levels are protective. Serum from 23 cases of malignant melanoma that developed between 1975 and 1987 in donors to the Washington County, Maryland serum bank was assayed along with that from 46 matched controls. There were no demonstrable differences among controls by sex, age, smoking or use of multivitamin supplements. The mean serum levels of 1,25-dihydroxyvitamin D were only 5% lower among cases than controls, a difference that could easily have occurred by chance.