alpha-Difluoromethylornithine is an enzyme-activated irreversible inhibitor of ornithine decarboxylase that forms a covalent bond with the enzyme. When alpha-[5-14C]difluoromethylornithine was administered to androgen-treated mice, only ornithine decarboxylase became labeled. Autoradiographic examination of kidney, liver, and brain indicated much more extensive incorporation of labeled difluoromethylornithine into kidney protein than into the protein of the other tissues. Such incorporation was greatly reduced by prior treatment of the mice with cycloheximide. These results correlate with the presence of ornithine decarboxylase activity which is much higher in the kidney than in the other tissues and is lost rapidly when protein synthesis is inhibited. The binding of this drug in vivo, therefore, is useful for determining the distribution of ornithine decarboxylase. The enzyme was predominantly located in the proximal tubule cells of the kidney in androgen-treated mice.