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The most commonly used tissue fixatives are aldehydes, such as formalin, which work by protein crosss-linking. Proper fixation of a specimen prevents antigen from being washed out and is a critical aspect of successful IHC staining. Cross-linking can mask antigens, preventing primary antibodies from binding, thus yielding weak staining or false negative results. Immunoglobulins are especially susceptible to formalin fixation.
Treatment with a proteolytic enzyme is required to digest excess aldehyde linkages and expose the antigen. HistoReveal is a proteolytic enzyme solution that can be used in place of traditionally used enzyme pretreatments, such as pepsin or trypsin, and results in superior staining. Unlike other enzymes, HistoReveal is gentle on tissue and does not destroy morphology.
Moreover HistoReveal requires only a brief incubation at room temperature. Performing antigen recovery with HistoReveal will allow you to dilute the primary antibody 5 to 10 fold further and achieve optimal staining.
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