Growing dogs were fed diets containing soy oil or poultry fat as the main fat source and soybean meal or meat meal as the main protein source to examine the effects of types of dietary fat and protein on fatty acid concentrations in serum and skin and on serum cholesterol concentrations. Dogs fed diets containing soy oil had higher serum linoleic acid concentrations and lower serum oleic acid, arachidonic acid, and cholesterol concentrations than dogs fed diets containing poultry fat. The type of dietary protein had marginal effects on fatty acid concentrations and did not affect serum cholesterol. Similar differences were found in cutaneous fatty acid concentrations, with soy oil-fed dogs having significantly (P < 0.05) higher linoleic acid and lower oleic acid concentrations in their skin than had poultry fat-fed dogs. This study suggested that dietary fat source influences serum and cutaneous fatty acid concentrations and serum cholesterol concentrations in dogs, irrespective of dietary protein source.