The purpose of this study was to evaluate cyclooxygenase (COX)‐1 and ‐2 activation in response to an acute bout of resistance exercise (RE) in human skeletal muscle. Before, 4, and 24 hours after a bout of unilateral knee extensor RE (3 sets of 10 repetitions at 70% of maximum) muscle biopsies were taken from the vastus lateralis of seven young males (25±0.4 y). Biopsy samples were assayed for COX‐1 and COX‐2 activity (700200, Cayman Chemical). COX‐1 activity was not increased with RE (PRE: 5.0±1.0, 4 hrs: 6.5±1.1, 24 hrs: 7.7±1.9 nmol PGH2·g muscle−1·min−1) while COX‐2 activity was elevated at 24 (p<0.05) but not four hours after exercise (PRE: 4.5±1.1, 4 hrs: 5.8±1.1, 24 hrs: 8.9±1.4 nmol PGH2·g muscle−1·min−1). Prostaglandin (PG) production due to COX‐1 and COX‐2 was similar (COX‐1/COX‐2 ratio: 1.1±0.3) before RE. This ratio (0.8±0.3) decreased at 24 hours indicating a greater proportion of PG production from COX‐2. COX activity has been implicated as a necessary component of a RE‐induced increase in skeletal muscle protein synthesis. Muscle protein synthesis is elevated as early as 1–4 hours after RE. Thus our data suggest that an elevation in COX‐1 or COX‐2 activity may not be necessary for the RE‐induced increase in protein synthesis. Our findings also support the general consensus that COX‐2 is inducible and that COX‐2 enzyme activity is present in resting skeletal muscle.Funding: MWU Start‐Up Funds, and MWU College of Health Sciences