PRODUCTION OF POLYCLONAL ANTIBODIES AGAINST PROTEINS AND PEPTIDES

This chapter focuses on the most commonly used procedures for production, purification, and characterization of polyclonal antibodies against proteins and peptides raised in rabbits and mice. The main purpose of the immune system is to bind and eliminate foreign molecules (antigens), such as bacterial toxins that have invaded an individual. This task is mainly accomplished by immunoglobulins, which are water-soluble glycoproteins that carry the specific antibody activity. The main reason for the preferential use of polyclonal antibodies is that their generation is rather simple, quick, and inexpensive, whereas the production of monoclonal antibodies requires equipment and skills in cell culture techniques and is relatively time consuming and expensive. In addition, a suitable test system must be established to screen hundreds of cell culture supernatants to identify and select the clones—producing the monoclonal antibodies of interest. Polyclonal antibodies raised against a protein are usually directed against more than one epitope. An advantage of polyepitopic specificity is that denaturation of proteins by SDS or by fixation with aldehydes usually will not destroy all epitopes so that polyclonal antibodies can be used in most instances for both immunoblotting and for immunostaining of chemically fixed cells and tissues. © 1993, Academic Press Inc.