Angiopoietin-like protein 4 (ANGPTL4; also known as fasting-induced adipose factor) is a plasma protein that stimulates oxidation of fatty acids and inhibits fat accumulation. The gastrointestinal tract appears to play an important role in regulating plasma ANGPTL4 concentration in some situations and may be influenced by microbes within the gastrointestinal tract. Our aim was to determine which tissues express ANGPTL4 in the bovine. Rumen, omasum, abomasum, duodenum, jejunum, ileum, colon, pancreas, liver, and subcutaneous adipose tissue samples were collected postmortem from 2 steers. Abundance of ANGPTL4 messenger RNA was quantified by quantitative real-time PCR, and was most abundant in liver and adipose tissue (P < 0.05). We also detected ANGPTL4 messenger RNA throughout the gastrointestinal tract, although its abundance was approximately 10% of that found in liver and adipose tissue. Western blot analysis revealed that ANGPTL4 protein was most abundant in liver and adipose tissue (P < 0.05), but omasal, abomasal, and ileal samples contained at least 60% as much ANGPTL4 protein as the liver and adipose tissue samples, and the protein was detected in all tissues. Finally, cross-sections of the liver, pancreas, and rumen wall were used for indirect immunofluorescent detection of ANGPTL4. Despite the low abundance of ANGPTL4 measured by quantitative real-time PCR and Western blot in ruminal tissue, immunofluorescence demonstrated that expression of ANGPTL4 in ruminal epithelial cells was equivalent to or greater than that in liver hepatocytes. These findings indicate that, as in other species studied, liver and adipose tissue are key sources of ANGPTL4 in cattle. However, the protein was also highly abundant in ruminal epithelium, making it possible that commensal microbes may influence ANGPTL4 synthesis and secretion in the ruminant gastrointestinal tract.