Development of Protocol for Production of Primary Antibody against Ovalbumin Protein in Chicken for Detection of the Protein through Western Blotting

ABSTRACT

Ovalbumin, a major protein of egg white plays many roles including providing nutrition to the developing embryo, acting as coagulating agent, folliculogenesis and angiogenesis in chicken and other animals. This protein is expressed mainly in magnum and then deposited over the yolk of the oocyte/zygote. Hence, it is important in formation of egg and is an essential target to measure. We cloned chicken ovalbumin CDS in pAcGFP-C1 vector and has been initially expressed in chicken primary magnum cell culture. The ovalbumin protein tagged with 6x Histidine was purified from cell culture and used for production of primary antibody in rat. The ovalbumin protein along with freund’s adjuvant was injected to the rat, booster was given, and finally, hyper-immune sera was collected from rat. The antisera was purified for isolation of IgG. The IgG was used as primary antibody for Western blotting. Through Western blotting, ovalbumin protein isolated from chicken magnum was detected and the protocol was established to detect chicken ovalbumin protein.