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The page below is a sample from the LabCE course Immunohistochemistry (IHC) Basics in Histology. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Secondary Antibody

If using a procedure (such as the ABC method) that requires a secondary antibody, the secondary is usually biotinylated or is conjugated with a fluorophore or chromogen. The secondary antibody must be an antibody against the primary antibody’s host.
Example: If the primary antibody is made in mouse the secondary or linking antibody should be horse anti-mouse.
The secondary antibody is also referred to as the linking antibody, where its purpose is to add a place for attachment of the avidin-biotin enzyme complex. In an immunofluorescence staining procedure, it is the last step if the secondary is conjugated with a fluorophore.