Endogenous Biotin Staining as an Artifact of Antigen Retrieval with Automated Immunostaining

Authors: Amitabh Srivastava; Arthur S. Tischler; Ronald A. Delellis

Source: Endocrine Pathology, Volume 15, Number 2, June 2004 , pp. 175-178(4)

Publisher: Humana Press

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Abstract:

We wish to highlight certain artifacts of antigen retrieval that we encountered recently while using automated immunohistochemistry. Immunohistochemical analysis for routine diagnostic purposes is increasingly being performed on automated stainers. The recognition of spurious immunoreactivity in such procedures is critically dependent on proper negative controls run with every batch of staining. Negative controls used most often involve either the omission of primary antibody or replacement of the primary antibody with normal serum at equivalent concentration. While most manufacturers provide appropriate negative control antisera in the form of irrelevant mouse and rabbit IgG to be used in place of the primary antibody, the role of antigen retrieval in negative controls is seldom highlighted. The phenomenon of heat induced endogenous biotin activity has been described previously [1– 3] and can be a pitfall in the interpretation of immunohistochemical results.

Document Type: Miscellaneous

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