Oxidative stress causes biochemical and morphological alterations in erythrocytes. The primary factors contributing to oxidative stress are aging and storage. Antioxidants significantly alleviate oxidative stress. Therefore, this study aimed to investigate the response of young and old erythrocytes to Vitamin C and Vitamin E during storage. Erythrocytes were separated into young and old by the Percoll method. Each erythrocyte subpopulation was categorized into i) Control [Additive Solution-7 (AS-7)] and ii) VC+VE group [Vitamin C and Vitamin E in AS-7] and stored for 21 days at 4˚C. Oxidative stress, antioxidant, and aging markers were analyzed on days 1, 14, and 21. The activity of antioxidant enzymes was similar throughout storage in Young cells. However, superoxide dismutase activity elevated in Old cells (Control and VC+VE) on days 1 and 21. Catalase activity increased on days 14 and 21, while glutathione peroxidase increased on days 1 and 14 in Old Controls. However, in Old VC+VE, catalase increased on day 21 and glutathione peroxidase increased on day 1. Advanced oxidation protein products, superoxides, glutathione, and uric acid increased in old cells throughout the storage. Malondialdehyde decreased in Old VC+VE compared to Old Control on days 14 and 21 respectively. Sialic acids and glutamate oxaloacetate transaminase activity were higher in young cells compared to old cells. Young cells exhibited lower oxidative changes throughout storage. Vitamin C and Vitamin E were effective in maintaining the redox balance in old cells. These findings emphasize the need for specific approaches for different subpopulations during erythrocyte banking.