Immunofluorescence detection of occludin in rat testicular tissue
Immunofluorescence is an antigen-antibody reaction in which the antibodies are labeled with fluorescent dyes and the formed antigen-antibody complex is visualized using fluorescent microscope. Two types of immunofluorescence assay are present, direct assay in which the antigen is allowed to bind with specific (primary) antibody that is labeled with a fluorescent dye, while in the indirect assay; the antigen is allowed to react with unlabeled primary antibody which in turn reacts with a secondary labeled antibody. This secondary antibody binds to Fc portion of the primary antibody. On washing, the unbound antibodies get washed off, while the bound antibodies remain (Odell & Cook, 2013). In the present study, the indirect immunofluorescence assay was adopted.