An experiment was conducted to determine the effect of the source of fat (soybean oil or tallow) on the ileal endogenous amino acid (EAA) losses in broilers. Three nitrogen (N)-free diets; a control diet with no added fat and test diets with 60 g/kg of either soybean oil or tallow were formulated. Titanium dioxide (5 g/kg) was added to all diets as an indigestible marker. Each diet was assigned to six replicate cages (eight birds per cage) from d 18 to 21 post-hatch. On d 21, the digesta were collected from the lower half of the ileum. The endogenous losses of nitrogen and amino acids (AA) were lower (p = 0.08; p = 0.001) in broilers fed diets with soybean oil or tallow, respectively, compared to those fed the diet with no fat. Source of fat had no influence (p > 0.05) on EAA losses. The most abundant AA in the ileal endogenous protein was glutamic acid, followed by aspartic acid, threonine, leucine, serine, valine and proline. In general, the concentrations of AA in the endogenous protein were lower (p < 0.05) with added fat. The exceptions were methionine, cysteine, proline and serine, which were unaffected. The effect of fat source on the AA contents of endogenous protein were inconsistent and differed depending on the AA.