Double Immunoenzyme Staining Methods with Special Reference to Monoclonal Antibodies.

Abstract

Immunohistochemical methods have been developed to evaluate the distribution patterns of different antigens (e.g. each characteristic for a certain cell type) simultaneously in tissue sections. These methods have relevance, especially for research in (tumor) cell biology. The demonstration of co-expression of cytoskeletal proteins in a single cell or the presence of different hormones in adjacent cells are good examples of such application. In the past double immunofluorescence techniques have been most widely used for this purpose. Double immunoenzyme staining however, is in many cases more attractive than double immunofluorescence because the preparations are permanent and the evaluation of the staining in relation to morphology is simple. Although several methods for double immunoenzyme staining have been published, their execution often remains problematic, especially when monoclonal antibodies (MoAb’s) are used. In this chapter we will review some of these methods and pay special attention to their practical applicability. Moreover, a method for double staining with MoAb’s will be described. For the common immunocytochemical techniques, the reader is referred to chapter 2.