Highlights • AX was determined in wheat grain after commercial and laboratory alcohol production. • Samples from laboratory scale production were similar to those from commercial processing. • The concentration, solubility and dynamic viscosity of AX increased during processing. • AX in DDGS was less variable in composition and properties than that in whole grain. • DDGS has potential to provide a source of AX for novel products. Laboratory produced DDGS samples were compared with commercial samples from a distillery and a biofuel plant. Changes in structure, solubility and content of arabinoxylan (AX) was determined. The distillation process results in a relative increase of AX content compared to the starting material. The heating and drying processes involved in the production of DDGS lead to an increased solubility and viscosity of water-extractable AX. Production of DDGS results in structural changes to the AX. There is a decrease in 2- and 3-linked arabinose oligosaccharides, that contributes to around a 50% reduction in arabinosylation in DDGS compared with the starting grains. The current study shows that laboratory-scale DDGS provide an accurate representation of the commercial scale and that the AX composition of DDGS is consistently uniform irrespective of starting material. The uniformity of DDGS and thin stillage makes them a good potential source of AX for production of prebiotics or other novel products.