We hypothesized that, at a common age endpoint, adipose tissue from corn-fed steers would be less sensitive to insulin than adipose tissue from hay-fed steers. Angus steers were assigned to either a corn-based diet (n = 6) or hay-based diet (n = 6) and fed to common days on feed. Steers fed the corn-based diet had 2.44 cm of fat thickness over the 12th thoracic rib, whereas hay-fed steers had 1.04 cm of fat thickness. At slaughter, subcutaneous adipose samples were collected and portions of subcutaneous adipose tissue were incubated with [U-(14)C]acetate to quantify fatty acid synthesis or with [U-(14)C]glucose to assess glucose utilization in the presence of 0, 100, or 500 ng/mL of insulin. Additional subcutaneous samples were used to evaluate glycolytic intermediate concentrations as indicators of glycolytic flux. Data were analyzed as a split-plot with diet in the main plot and insulin concentration and its interaction with diet in the sub-plot. Within diet, linear and quadratic contrasts of insulin concentration were tested. Diet had no effect (P > or = 0.31) on glucose metabolism or acetate carbon incorporation into total lipids (P = 0.32). Insulin had no effect (P > 0.21) on glucose conversion to CO(2), lactate, or total lipids, nor did it affect (P = 0.28) acetate conversion to total lipids. No diet x insulin interaction (P > 0.36) was observed for any measure of subcutaneous metabolism in vitro. Steers fed the corn-based diet exhibited neither a linear (P > 0.22) nor a quadratic (P > 0.24) effect to increasing insulin concentration. However, when steers were fed the hay-based diet, there was a positive linear (P = 0.06) effect for glucose oxidation. These results suggest that subcutaneous adipose tissue may become resistant to stimulation by insulin in steers fed to a fat thickness above the average feedlot steer, but this is independent of diet.