A Monoclonal Antibody Against Bovine Adiponectin

Abstract

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.

Characterization of adiponectin concentrations and molecular weight forms in serum, seminal plasma, and ovarian follicular fluid from cattle

“…In the serum, the MMW was the primary band, whereas only a faint band for the HMW was observed. Similar MW patterns of the AdipoQ protein in serum were shown for dairy cows [24][25][26]. In contrast to the serum, the SP exhibited a prominent HMW band in addition to the MMW band, which was also present.…”