Indirect Immunofluorescence of Cultured Cells

Abstract Immunofluorescence is an immunoassay for the detection of specific cellular constituents using antibodies conjugated to a fluorochrome. Indirect immunofluorescence refers to a two‐step procedure, whereby the target‐specific, primary antibodies are applied unlabelled and are detected with fluorochrome‐conjugated secondary antibodies. Its main applications are identification and localization of cell components by fluorescence microscopy, and automated identification, sorting and quantification of fluorescent cells by flow cytometry.