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The objective of this study was to evaluate the effect of timing of electrical stimulation on free calcium concentration, calpain-2 activity, Warner-Bratzler shear force (WBSF), and consumer sensory analysis. Twenty-three beef steers were harvested and stimulated (S) using extra-low voltage or not stimulated (NS), at exsanguination and/or 1 h postmortem, resulting in 4 different stimulation treatments: NS-NS, NS-S, S-NS, or S-S. Samples were removed from the longissimus lumborum (LL) and semimembranosus (SM) for free calcium and calpain-2 analysis on days 1, 4, and 14 postmortem. WBSF and sensory analysis steaks were removed on day 4 and frozen (4 d) or aged to 14 d postmortem. Data were analyzed using the mixed model or generalized linear mixed model procedure of SAS (SAS Institute, Inc., Cary, NC), with significance determined at P < 0.05. There was a tendency for an aging-period-by-stimulation-treatment interaction for LL free calcium concentration (P = 0.05), and there was a significant difference between aging periods (P < 0.01). No difference was observed in free calcium concentration in the SM between stimulation treatments (P = 0.44); aging, however, significantly increased SM free calcium concentration (P < 0.01). Stimulation did not impact native calpain-2 activity in the LL (P = 0.71) or SM (P = 0.89). Stimulation treatment did not improve tenderness values for WBSF analysis for the LL (P = 0.69) or SM (P = 0.61) or consumer sensory analysis in the LL (P = 0.56) or SM (P = 0.36). A longer aging period tended to increase calpain-2 activity in the SM (P = 0.08), improve WBSF in the LL (P = 0.09), and significantly improve consumer tenderness scores in the SM (P < 0.01). In conclusion, the timing of electrical stimulation utilized in the current study tended to influence free calcium concentration in the LL but did not influence calpain-2 activity or beef tenderness. Aging, however, improved tenderness.