Adiponectin (AdipoQ) is an adipokine mainly secreted by white fatty tissue, playing a major role in energy homeostasis and insulin sensitivity. For cattle, AdipoQ data are largely limited to mRNA expression; to our knowledge, valid information about the AdipoQ protein in bovine tissues and body fluids is not available. Therefore, we have developed a monoclonal antibody against bovine AdipoQ. This study describes the preparation, application, and characterization of a monoclonal antibody for use in ELISA, Western blot, and histology. The antibody was developed by PEG fusion of the SP2/0 cell line with splenic B cells from AdipoQ immunized C57Bl/6 mice. Antibody-producing cells were identified by ELISA and specified by immunoblotting and immunostaining of bovine retroperitoneal adipose tissue. The novel antibody detects AdipoQ in histological samples, ELISA, and Western blots.