Abstract. Feeding wet distillers grains plus solubles (WDGS) increases polyunsaturated fatty acid (PUFA) levels in beef. It was hypothesised that WDGS in feedlot diets increases PUFA concentration in the sarcoplasmic reticulum (SR) membrane, thereby altering membrane integrity, resulting in more rapid intracellular calcium leakage and improved tenderness. The objective of this study was to evaluate this hypothesis. Ninety-six crossbred steers were fed either a corn-based diet with 0% WDGS or 50% WDGS. Fifteen strip loins per treatment were collected, fabricated into steaks, aged and placed under retail display conditions. Steaks were used to measure tenderness, proteolysis, free calcium concentrations, lipid oxidation, sarcomere length and SR membrane fatty acid, phospholipid lipid, neutral lipid and total lipid profiles. Compared with steaks from steers fed 0% WDGS, steaks from steers fed 50% WDGS were more tender (P < 0.05) and had greater (P < 0.05) free calcium concentrations early post-mortem. Feeding 50% WDGS also tended to increase (P < 0.10) total PUFA concentrations, decrease (P < 0.10) total phospholipid concentration and increase (P < 0.10) total neutral lipid concentration for SR membrane. Steaks from steers fed 0% WDGS had greater (P < 0.05) lipid oxidation (TBARS values) than steaks from steers fed 50% WDGS after extended aging. Although differences in tenderness between the two treatments were detected, there were no corresponding differences (P > 0.10) in sarcomere length or proteolysis. This study showed that feeding WDGS may increase tenderness, possibly by increasing free calcium in muscle early post-mortem. However, the true mechanism that contributes to these differences is still unclear.