A solid-phase beta-galactosidase ELISA for detecting and quantifying monoclonal antibody binding to dissociated cell cultures of postnatal rodent cerebellum.
A solid-phase, indirect beta-galactosidase-linked immunoassay (ELISA) is described for screening large numbers of monoclonal antibodies that recognize cell surface antigens of primary monolayer cerebellar cultures. Target cultures were prepared from perikaryal suspensions of postnatal rodent cerebellum seeded into poly-L-lysine pre-coated, flat-bottom microtiter wells and fixed with glutaraldehyde after growth in vitro. Hybridoma supernatants were then incubated on these cultures. After the addition of beta-galactosidase-linked anti-mouse IgG F(ab')2 fragments, antigen-positive supernatants were detected with the enzyme substrate o-nitrophenyl-beta-D-galactopyranoside. Using a monoclonal antibody specific for rat brain Thy-1 glycoprotein, this solid-phase ELISA was found to be useful in quantifying changes in the developmental expression of cerebellar surface antigens in these cultures.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D001666:Binding Sites, Antibody; D002531:Cerebellum / Q000276:immunology*; D046508:Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005696:Galactosidases; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D051379:Mice; D008808:Mice, Inbred CBA; D001616:beta-Galactosidase
D000911:Antibodies, Monoclonal; D005696:Galactosidases; D001616:beta-Galactosidase