ABSTRACT
Lipopolysaccharide (LPS) is the major carbohydrate component of Gram-negative bacteria, and forms an integral part of the outer membrane. This chapter describes a procedure for the preparation of LPS based on the hot-phenol method of O. Westphal and K. Jahn and also a modification of the hot-phenol procedure developed for obtaining LPS from bacteria which have strong associations between LPS and outer-membrane proteins. It also describes the use of SDS-PAGE for the characterization of purified and whole-cell LPS. Because of the highly antigenic nature of LPS and the importance of antibody reactions with the O-antigen, the use of LPS in immunoblotting for reaction with antibodies and a simple method for preparing LPS for producing antisera will be discussed. The method of LPS purification is a modification of the hot-phenol procedure of Westphal and Jahn. The amount of LPS preparation used for antiserum production will depend on the species and size of the animal of choice.
