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.