We estimated ruminal crude protein degradation of twelve feedstuffs commonly used in China using in vitro and in vivo methods. The in vivo net protein utilization (NPU) levels of corn, sorghum, barley, wheat, Chinese wild rye grass, corn stalk, rice straw, soybean straw, soybean meal, distillers' dried grains with solubles (DDGS), Brewers' spent grains, and sunflower meal were 52.57, 49.68, 65.38, 72.58, 82.41, 72.26, 68.57, 76.95, 54.75, 56.27, 29.03 and 41.88%, respectively. The linear regression between NH3-N incorporated into microbial proteins and gas production after incubation (6, 12, and 24 h) was significant (r = 0.9948 and P < 0.001, r = 0.9874 and P < 0.01, and r = 0.9912 and P < 0.01, respectively). Based on the linear regression equations, we estimated in vitro protein degradability (IVPD) and generated the regression equations between IVPD and NPU. The linear regression equations between IVPD and NPU after 6 h incubation in the energy, protein, and roughage feed groups were Y = 0.5633X + 33.20 (R2 = 0.8517, P < 0.05), Y = 0.8482X+ 34.81 (R2 = 0.8650, P < 0.05), and Y = 1.6295X - 17.70 (R2 = 0.909, P < 0.05), respectively. The in vitro gas production method is useful for the determination of protein degradation in feedstuffs.