In order to integrate and analyze knowledge on the use of protein supplements and protein nutrition of lactating dairy cows, we compiled a review of 108 studies published throughout the world, but principally in the Journal of Dairy Science between 1985 and 1997. In 29 comparisons from 15 metabolism trials, soybean meal was replaced by high amounts of rumen undegradable protein (RUP) as a supplement; the benefits were not consistently observed for flow to the duodenum, essential amino acids, or lysine and methionine. High RUP diets resulted in decreased microbial protein synthesis in 76% of the comparisons. However, fish meal provided a good balance of lysine and methionine when calculated as a percentage of total essential amino acids. In 127 comparisons from 88 lactation trials that were published from 1985 to 1997, researchers studied the effects of replacing soybean meal with high RUP sources, such as heated and chemically treated soybean meal, corn gluten meal, distillers grains, brewers grains, blood meal, meat and bone meal, feather meal, or blends of these sources; milk yield was significantly higher in only 17% of the comparisons. Fish meal and treated soybean meal accounted for most of the positive effects on milk yield from RUP; corn gluten meal resulted in mostly negative results. The percentage of fat in milk was depressed more by fish meal than by other RUP sources. Protein percentage was decreased in 28 comparisons and increased in only 6 comparisons, probably reflecting the decrease in microbial protein synthesis, as was observed for diets high in RUP. The data strongly suggest that increased RUP per se in dairy cow diets, which often results in a decrease in RDP and a change in absorbed AA profiles, does not consistently improve lactational performance.