Preparation of polyclonal antibodies with recombinant N terminus of porcine lactotransferrin

Porcine lactotransferrin(LTF) was found to be a potential receptor protein during screening receptors that reacted with MSG1 of by affinity chromatography.In order to further confirm the receptor function,the truncated LTF was expressed and polyclonal antibodies were prepared.Firstly,a 762 bp fragment at the N-terminus of porcine lactotransferrin was amplified from synthesized full-length gene of porcine LTF and cloned into the plasmid pET-32a(+).Secondly,the recombinant plasmid was transformed into Escherichia coli competent cells BL-21 and induced by IPTG.The recombinant protein was identified specifically by His monoclonal antibody and purified by inclusion body lotion.Finally,mice and rabbits polyclonal antibodies were prepared by back multi-intracutaneous injection with purified protein.The results showed that the recombinant protein was approximately 44 ku and specifically reacted with His monoclonal antibody.Western-blot result showed the mouse and rabbit polyclonal antibodies were specifically reacted with recombinant LTF.The preparation of mice and rabbits polyclonal antibodies will provide technical assist for the confirmation of porcine LTF receptor function and the location of porcine LTF by confocal laser technology.