Antibody Engineering for Pursuing a Healthier Future

Saeed, Abdullah F. U. H.; Wang, Rongzhi; Ling, Sumei; Wang, Shihua

Immunohistochemistry (IHC) is used for the detection and identification of proteins and their localization in tissues. It is essential to retain the morphology of tissues, cells and the availability of antigen sites. Fresh, rapidly frozen tissue sections are preferably used for IHC by chemically fixing tissues in formalin and embedding in wax (Zhu et al., 2015) . Additionally, fixing crosslinks amino acids in the tissue that block access to the epitope sites and prevent the action of any protein specific antibodies. The exposure of hidden epitope regions is performed by digestion with an enzyme or by heat treatment, which removes endogenous peroxidase activity and non-specific sites are blocked. A labeled antibody or an unlabeled primary antibody specific to the protein of interest is used, followed by the addition of a secondary labeled antibody specific to the primary antibody (Diorio et al., 2016) .