Abstract. The unavailability of homologous antibodies to avian thyrotropin (TSH) has been a major hindrance in the study of avian reproduction and metabolism. Here we report the production and characterization of an anti-peptide antibody specific to chicken TSH β subunit. According to the cDNA sequence of chicken TSH β subunit, a synthetic peptide corresponding to the amino acid residues 35-50 (Tb3550; Asp-Ser-Asn-Gly-Lys-Lys-Leu-Leu-Leu-Lys-Ser-Ala-Leu-Ser-Gln-Asn) was prepared to produce polyclonal antibody in rabbits. This peptide sequence is highly specific to avian TSH β and was designed for the antibody to show no cross-reaction to pituitary and other chicken proteins with known amino acid sequences. Anti-Tb3550 recognized a 18.2-kDa band on immunoblot of the cephalic lobe homogenate, but not of the caudal lobe homogenate, of chicken anterior pituitary gland. Positive immunohistochemical staining with anti-Tb3550 was restricted to the cephalic lobe, whereas the cells positive to anti-chicken luteinizing hormone were scattered throughout the anterior pituitary. Anti-Tb3550 did not bind either to purified chicken luteinizing hormone or to follicle stimulating hormone preparation. These results indicate that this antibody specifically recognizes chicken TSH β subunit. This antibody should be useful for immunochemical study of avian TSH and for tracing TSH activity during the purification of the hormone. Key words: Chicken, TSHβ, Anti-peptide antibody, Immunoblotting, Immunohistochemistry (J. Reprod. Dev. 48 : 197-204, 2002) vian thyroid hormones have been reported to affect not only heat production, metabolism and growth as in mammals, but also gonadal development, molting and migration (see [1][2][3] for review). The involvement of thyroid hormones in induced molt of the hen, where hens are forced into molting to improve egg production and egg quality, in particular has attracted the attention of investigators due to its biological interest as well as economic importance to the poultry industry [4][5][6]. Whether the natural seasonal molt is brought about by increased thyroid activity seems less certain, but t h e r e h a v e b e e n r e p o r t s s u g g e s t i n g t h e involvement of the thyroid in wild avian species (see [7] for review). Although thyrotropin (TSH), as a tropic hormone of the pituitary gland, is supposed to regulate circulating thyroid hormone levels under various physiological and artificial conditions such as molting, the exact roles of TSH are not well understood, since purification with thorough characterization of avian TSH has not been successful and hence no homologous immunoassays specific for avian TSH are yet available.