This study was designed to compare the fatty acid profiles of four types of fat depots from bulls, steers, and heifers. All animals were Polish Holstein‐Friesian and Limousin crossbreds fattened semi‐intensively—silage at libitum and concentrate in the amount corresponding to 30% of their net energy requirements. The fatty acid profile in intramuscular, intermuscular, and external and internal fat was determined. The intramuscular fat of bulls was most abundant in total PUFAs and n‐6 PUFAs, and functional fatty acids C 18:2, C 20:4, and C 22:5 in comparison with steers and heifers. Regardless of sex category, intramuscular and external fat were characterized by higher levels of UFAs and a more desirable MUFA/SFA ratio than the remaining fat types. It should also be noted that external fat was more abundant in CLA than other fat types, and that the highest CLA content was found in bull fat, compared with the remaining sex categories of cattle.