| Resumo: |
Protocols for blocking non-specific antibody (Ab) binding in immunohistochemistry are based on rather contradictory and outdated reports. This prompted us to prove, whether non-specific Ab binding may really lead to unwanted background staining in routinely processed cell and tissue probes. In this study, the probes were fixed and processed according to routine protocols with and without a blocking step (goat serum or BSA). Surprisingly, all Ab in probes processed without a blocking step did not show any propensity towards non-specific binding that might lead to background staining, thus implying that endogenous Fc receptors do not retain their ability to bind Fc portion of Ab after standard fixation. Likewise in routinely fixed probes, we did not find any non-specific Ab binding ascribed to a combination of ionic and hydrophobic interactions. The traditionally used protein blocking step is useless in immunostaining of routinely fixed tissues.
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