Indigestible neutral detergent fiber (NDF) predicts forage digestibility accurately and precisely when determined by a 288-h ruminal in situ incubation, and it is an important parameter in mechanistic rumen models. The long incubation time required is a disadvantage. Further, intrinsic cell wall characteristics of feeds should be determined under ideal conditions for fiber digestion. The objective of this study was to determine the effects of diet composition and rumen incubation time on the concentrations of indigestible NDF (iNDF) for a wide range of feeds in dairy cows. Additionally, predicted concentrations of unavailable NDF generated using the National Research Council (NRC) model and the Cornell Net Carbohydrate and Protein System (CNCPS) were evaluated. Indigestible NDF was evaluated in 18 feeds using 4 cows in a split-split plot design. Treatments were in a 3 × 3 factorial arrangement, consisting of different diets and incubation times. Diet composition was primarily varied by changing the level of concentrate supplementation between 190 (low), 421 (medium), and 625 (high)g/kg of diet dry matter (DM). Grass silage was used as the basal forage for all cows. The feeds were incubated for 144, 216, and 288 h. Indigestible NDF was determined from 2-g samples weighed into polyester bags with a pore size of 12 µm and a pore area equal to 6% of the total surface area, giving a sample size to surface ratio of 10mg/cm(2). Across all feeds, the measured iNDF concentrations ranged from 6 to 516 g/kg of DM. The feed iNDF concentration was not affected by the cow used, but diet composition had a significant effect. The mean measured iNDF concentrations for cows consuming low-, medium-, and high-concentrate diets were 178, 186, and 197 g/kg of DM, respectively. The incubation time also affected the feed iNDF concentrations, which averaged 199, 185, and 177 g/kg of DM for 144-, 216-, and 288-h incubations, respectively. We also observed significant interactions between incubation time and feed, and between diet composition and feed, with fiber-rich feeds being most sensitive to these factors. The evaluation of model predictions of unavailable NDF indicated poor precision with prediction errors of 56 (NRC) and 84 (CNCPS)g/kg of DM. Indigestible NDF should be determined based on 288-h ruminal in situ incubations in cows consuming diets with a low proportion of concentrate to represent the feed fraction that is unavailable to the animal.