Quantitative protein profiling of milk of Nili-Ravi buffaloes, Sahiwal and cross-bred cows were performed. Total proteins and caseins were the highest in milk of cross-bred cows while, whey proteins were the maximum in Sahiwal cow’s milk. The mean values of milk total proteins were 78.103±1.878, 59.333±0.611 and 81.594±2.026 mg/mL for Nili-Ravi buffaloes, Sahiwal cows and cross-bred cows, respectively. The milk lactoserum proteins were comparatively higher in milk of Sahiwal cows (28.842±1.25mg/mL) followed by Nili-Ravi buffaloes (20.026±0.665mg/mL), and the lowest in cross-bred cow’s milk (10.176±0.568mg/mL). The mean value of caseins in milk of Nili-Ravi buffaloes was58.077±0.90mg/mL, while the least (30.491±1.15mg/mL) in Sahiwal cow’s milk and the highest (71.418±1.20mg/mL) in milk of cross-bred cows. Effect of different quarters on milk protein profiles was non-significant in all three species/cattle breeds. Bulk milk contained 73.555±0.887 mg/mL total proteins, 19.586±0.767 mg/mL lactoserum proteins and 53.969±1.464 mg/mL caseins.