Fatty acid-binding proteins are considered to be the carriers for the transportation of intracellular fatty acids and play an important role in the development of fatness traits. Adipocyte fatty acid-binding protein (A-FABP) is one of the family members. The current study was designed to analyze the tissue expression characterization of chicken A-FABP and its expression difference between the fat and lean males in abdominal fat tissue to reveal the possible relationship between the expression of A-FABP and abdominal fat tissue development and growth in chicken. First, fusion protein glutathione S-transferase/A-FABP was induced and purified, and then the antiserum containing specific polyclonal antibodies was obtained by immunizing healthy female rabbits using the purified fusion protein. Second, tissue expression characterization of A-FABP was investigated by Western blot. Finally, A-FABP expression difference in abdominal fat tissue between the fat and lean males was investigated by real-time reverse transcription-PCR and Western blot methods. The results showed that A-FABP expressed specifically in abdominal fat tissue and the mRNA expression level of A-FABP in fat males was lower than that of lean males at 2, 3, 4, 6, 7, 9, and 10 wk of age (P<0.05), and the protein expression level of fat males was lower than that of lean males at 6 and 10 wk of age (P<0.05). These results suggested that chicken A-FABP might affect abdominal fat deposition through changing its expression level, and the possible mechanism may be that a high expression level of A-FABP induced the high lipolytic rate and led to the decreased abdominal fat mass accordingly.