FLUORESCENT MICROTITER SCREENING ASSAY FOR IMMUNOCYTOCHEMICALLY REACTIVE ANTIBODIES

We have developed a rapid in situ screening procedure that enables pre -screening of hundreds of hybridomas in a 96-well format. The procedur e involves fluorescence immunostaining of cells cultured in 96-well pl ates and the use of a fluorescence plate reader to detect reactive ant ibodies. Positive immunostaining in individual wells, as denoted by el evated readings, is then confirmed by fluorescence microscopy. Using t he method described here, we have successfully identified monoclonal a ntibodies that are specific to the nuclear receptor, peroxisome prolif erator-activated receptor gamma (PPAR gamma). This assay is readily ap plicable for screening hybridomas raised against to aid in the initial identification of antibodies reactive in immunocytochemical procedure s.