Bioethanol production has led to the production of considerable quantities of different coproducts. Variation in nutrient profiles as well as nutrient availability among these coproducts may lead to an imbalance in the formulation of diets. The objectives of this study were to fractionate protein and carbohydrates by an in situ approach, to determine ruminal availability of nutrients for microbial protein synthesis and to determine protein availability to dairy cattle for three types of dried distiller's grains with solubles (DDGS; 100% wheat DDGS (WDDGS); DDGS blend1 (BDDGS1, corn to wheat ratio 30 : 70); DDGS blend2 (BDDGS2, corn to wheat ratio 50 : 50)) and for different batches within DDGS type using the 2010 DVE/OEB protein evaluation system. The results indicated that all DDGS types are quantitatively good sources of true protein digested and absorbed in the small intestine (DVE values; 177, 184 and 170 g/kg dry matter (DM) for WDDGS, BDDGS1 and BDDGS2, respectively). Rumen degraded protein balances (OEB) values were 159, 82, 65 g/kg DM in WDDGS, BDDGS1 and BDDGS2, respectively. Despite the differences in ruminal availability of nutrients among the different batches of DDGS, the DVE values only differed between the batches of BDDGS1 (194 v. 176 g/kg DM). In conclusion, when DDGS is included in the rations of dairy cattle, variation in its protein value due to factors such as DDGS batch should be taken into consideration.