Endogenous biotin staining as an artifact of antigen retrieval with automated immunostaining.

摘要

We wish to highlight certain artifacts ofantigen retrieval that we encounteredrecently while using automated immunohistochemistry.Immunohistochemicalanalysis for routine diagnostic purposes isincreasingly being performed on automatedstainers. The recognition of spuriousimmunoreactivity in such proceduresis critically dependent on proper negativecontrols run with every batch of staining.Negative controls used most often involveeither the omission of primary antibodyor replacement of the primary antibodywith normal serum at equivalent concentration.While most manufacturers provideappropriate negative control antisera in theform of irrelevant mouse and rabbit IgG tobe used in place of the primary antibody,the role of antigen retrieval in negative controlsis seldom highlighted. The phenomenonof heat induced endogenous biotinactivity has been described previously [1-3] and can be a pitfall in the interpretationof immunohistochemical results.