Effects of the β-agonist, ractopamine-HCl (ractopamine), on skeletal muscle protein turnover were evaluated in 16 steers (512 kg) and 16 heifers (473 kg). Treatments were arranged in a 2 × 2 factorial design and included gender (steer vs. heifer) and ractopamine (0 or 200 mg/d). Steers were implanted with 120 mg trenbolone acetate and 24 mg estradiol-17β, heifers with 140 mg trenbolone acetate and 14 mg estradiol-17β. Cattle were fed a diet based on steam-flaked maize. Muscle biopsy samples were collected from m. longissimus and m. biceps femoris on day 0 (prior to ractopamine feeding) and after 14 and 28 d of ractopamine feeding. In vitro protein synthesis rates in m. longissimus tended to be greater for steers than for heifers, but no effect of gender was observed in m. biceps femoris. Gender did not affect in vitro protein degradation rates for either muscle. Ractopamine significantly decreased rates of in vitro protein degradation in m. longissimus, but rates in m. biceps femoris were not affected by ractopamine. Ractopamine did not affect in vitro protein synthesis rates. In general, there were no striking differences between steers and heifers in response to ractopamine, indicating that its effectiveness should be similar between genders.