This research was performed to detect tissue fatty acid (FA) composition and conjugated linoleic acid (CLA) content in Anatolian buffaloes at different muscle types (MTs) and slaughter weights (SWs). The research was carried out on a private commercial livestock farm in Tokat. The research's animal material comprised 20 Anatolian buffalo calves with approximately 100 kg body weights, weaned at about 150 days of age. Before the experiment started, the calves were randomly divided into two slaughter groups (SW) as low weight (LW=225 kg) and high weight (HW=325 kg). Ten calves from each of the two experimental groups, which were intensively fed, were slaughtered in two different weights. After the slaughtering, FA composition and CLA content of the Semimembranosus (SM), Semitendinosus (ST), and Triceps brachii (TB) muscle tissues of the animals were examined. The study determined that palmitoleic acid (C16:1) and stearic acid (C18:0) were affected by MTs and oleic acid (C18:1) and alpha-linolenic acid (C18:3 n-3) were affected by SW (P < 0.05). The highest CLA was in the ST muscle type (0.298) and the LW group (0.289) of the SW groups (P > 0.05). With the increase of SW (in LW and HW groups), n-6/n3 (SM: 7.783 and 6.533; ST: 8.115 and 7.859; TB: 8.416 and 8.215) (P > 0.05) and PUFA ratio decreased (P < 0.05). The SW increase raised the SFA ratio in the SM muscle (P < 0.05) while lowering it in the TB muscle (P > 0.05). Again, with the increase in SW, AI and TI values increased in SM and ST muscles, while the same index values decreased in TB muscle (P> 0.05). In conclusion, when considering the PUFA/SFA ratio and the beneficial effects of CLAs on human health, ST in the MT and LW groups in SW, and thus ST and LW in MT and SW were prominent in Anatolian buffaloes.