Preparation of polyclonal antibody to CHOP protein and its application in heat stress of chickens.

Heat stress (HS) is a stress response of organisms to temperature changes, which can result in organ damage and increased chicken mortality in high-temperature environments. The CHOP protein, also known as GADD 135, plays a crucial role in endoplasmic reticulum stress. However, there are fewer studies related to whether CHOP proteins are involved in heat stress-induced organ damage. In this study, recombinant CHOP-pET-32a expression vector was constructed by using the prokaryotic expression technique of exogenous genes, and recombinant CHOP protein was obtained. Subsequently, rabbit anti-chicken CHOP polyclonal antibody was prepared by immunizing rabbits, and the antibody potency was higher than 1:102,400 as determined by ELISA. Immunofluorescence and western blotting demonstrated that the anti-CHOP antibody specifically recognized chicken CHOP protein. The protein was expressed in various organs, including the heart, liver, spleen, lung, kidney, bursa of Fabricius, and all segments of the intestine. Following heat stress, the expression of CHOP in the heart significantly increased, indicating a close association between CHOP and the occurrence of heat stress. The preparation of rabbit anti-CHOP polyclonal antibodies will be useful for future studies on poultry diseases.