Use of Multiple Antibodies for Labeling p53 Antigen

An antigen is a highly complex, three-dimensional molecule, and the precise location of epitopes on or within the antigen in most cases is not known. Furthermore, antigen retrieval methods may unfold the antigen molecule, exposing hidden (buried) epitopes. 

These methods may break down crosslinks between the antigen and the surrounding proteins, allowing the epitopes to become accessible due to the mechanism(s) responsible for the availability of epitopes for immunohistochemistry—a strategy to ensure that recognition of the epitope by the antibody is required. Such a protocol is presented below. 

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