Our objectives were to determine the effect of post rigor calcium chloride injection or freezing on 1) sarcoplasmic calcium concentration and calpain-2 activity of beef longissimus lumborum (LL) and semimembranosus (SM) steaks aged 1, 4, and 14 days post-treatment and on 2) Warner-Bratzler shear force, water holding capacity, and consumer acceptability of LL and SM steaks aged 4 and 14 days post-treatment. Free calcium levels in the calcium, frozen, and control steaks averaged 1256, 127, and 121 mu M for the LL and 1520, 120, and 111 mu M for the SM, respectively. Measurable LL native calpain-2 activity was lower in calcium and frozen steaks than control steaks (P < 0.01), while SM native calpain-2 activity was lowest in calcium steaks and intermediate in frozen steaks (P < 0.01). LL calcium steaks were more tender (P = 0.04) than control steaks. In conclusion, calcium chloride injection and freezing activate calpain-2 earlier postmortem in both muscles and calcium injection improves LL tenderness.