Four lactating dairy cows equipped with ruminal and duodenal cannulas were used to determine the impact of different methods of treating soybean meal (SBM) on the ruminal degradability and intestinal digestibility of crude protein and AA. Solvent-extracted SBM (SE), expeller SBM (EP), lignosulfonate SBM (LS), and heat and soyhulls SBM (HS) were incubated in the rumen in nylon bags for 48, 24, 16, 8, 4, 2, and 0 h according to National Research Council (2001) guidelines. Additional samples of each SBM product were also incubated for 16 h in the rumen; the residues from these bags were transferred to mobile bags, soaked in pepsin HCl, and then used for determination of intestinal digestibility in situ or in vitro. Treatment of SBM (EP, LS, HS) protected the crude protein and AA from ruminal degradation, increasing rumen undegradable protein from 42% in SE to 68% in EP. Kinetic analysis of crude protein and AA degradation in the rumen revealed that, compared with LS and HS, EP exhibited slower rates of degradation but a shorter lag phase and a higher proportion of soluble protein. For all SBM products, the pattern of ruminal degradation, at 16 h of incubation, was characterized by extensive degradation of Lys and His, whereas Met and the branched-chain AA were degraded to the least extent. Estimates of intestinal digestibility of AA and crude protein were lower when measured in vitro than in situ; the magnitude of the difference between the 2 methods was greater (25%) with treated SBM products than with SE (10%). The availability of essential and nonessential AA was consistently greater (30%) with treated SBM than with SE. Among the treated SBM products, 4 essential AA (Ile, Leu, Phe, and Val) showed differences in availability, with values consistently lower for HS than for LS. This study showed that, based on in situ measures, heat and chemical treatment of SBM enhanced AA availability, and that compared with HS, EP and LS had a higher potential to enhance the AA supply to the small intestine of high-producing dairy cows.