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Publisher Summary This chapter discusses the different aspects of the reverse plaque-forming cell (PFC) assay. The reverse PFC assay exploits the phenomenon of the reverse hemolysis wherein erythrocytes coated with purified antibody eventually lyse in the presence of the complementary antigen. It is found that when one incubates secretor cells, with erythrocytes, coated with antibody to the secreted molecules in an agar gel at 37°, the antigen molecules secreted by the cells diffuse outwardly and bind to the indicator erythrocytes to form antibody–antigen (Ab–Ag) complexes. These immune complexes usually do not bind enough complement to lyse the indicator cells efficiently. The specificity of the assay is dictated by the combined specificities of the indicator cells and of the developer. It is observed that, if the coating antibody is monospecific, the developing antibody may be used directly without any adsorption for specificity. The most popular use of the reverse PFC assay is the enumeration of the total population of Ig-secreting cells and of individual subpopulations of Ig isotype-, allotype-, and idiotype-secreting cells in mouse, in man, and in rabbit. |
