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The Avidin and Biotin solutions are color-coded with a pH indicator. In most cases, endogenous biotin in tissue sections is masked by formalin fixation. However, if avidin-biotin IHC detection systems are used with frozen sections or tissues pretreated with a heat-induced, epitope retrieval method (HIER), an avidin-biotin blocking technique may be needed. It can also be used on an automated staining system.