Abstract
In a variety of systems, secretion by individual cells, and changes of this function, which depend on cell adhesion/communication, can be studied using modifications of the original hemolytic plaque assay. This assay was developed for evaluating Ig production by lymphocytes (Jerne et al. 1974), and the modifications, referred to as reverse hemolytic plaque assays (RHPAs), are based on the complement-induced lysis of red blood cells (RBCs) bearing immunocomplexes. Practically, the cells of interest are attached to a support, together with RBCs that have been coated with staphylococcal protein A, and tested in the presence of polyclonal Ig raised against the secretory product (most often a protein) to be monitored. Binding of these antibodies to the product released by the cells is followed by the fixation of the immunocomplexes onto the surface of surrounding RBCs. After addition of complement, the immunocomplexes-bearing RBCs lyse, resulting in the formation of a hemolytic plaque around secreting cells (Fig. 12.1).
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© 1998 Springer-Verlag Berlin Heidelberg
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Bosco, D., Meda, P. (1998). Assessing Release of Secretory Products from Individual Cells. In: Clynes, M. (eds) Animal Cell Culture Techniques. Springer Lab Manual. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-80412-0_12
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DOI: https://doi.org/10.1007/978-3-642-80412-0_12
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