| | Re: Applications for antibodies
Otherwise from ELISA and WB methods, immunohistochemistry (IHC) it has the ability to show where certain proteins are located, inside a tissue section (normal or in relationship to its pathology), that deriving from a specific patient (age, sex, etc.), and effected biopsy at unfixed time. In IHC procedure, tissue section must be fixed before to prevent chemical or biological changes site of located proteins. Fixation process involves that proteins and other cellular components of the tissue, cannot stir anymore, to modify conformationally of it, to interact the one of it with the others, and this condition been termed “native molecules”. The produced antibodies against native molecules, in one conformational natural state (folded state), recognize only epitopes that has been present in this molecular form, while if the same proteins has been submitted to denaturation to get a linear molecules, such as WB during the preparation gel electrophoresis, the elicites antibodies cannot be able to recognize the same epitopes expressed on the native protein, still more if these are found in the fold (epitopic inaccessibility). Nevertheless, possibility that slightly denaturing tissue can to determine changes of native molecules with epitopes expression which are able to react with the antibody that detect same antigen in ELISA and WB methods. In add, two types of monospecific first antibodies to used in immunoassay : a) clonal, and b) monoclonal. The first recognize solely very specifically selected linear epitope on the antigen molecule after its detailed proteomic analysis, whereas the monoclonal antibodies recognize very often steric epitopes that frequently change their conformation during tissue preparation (denaturation tissue, protein extraction, protein transfer in the case of western blot – related procedures, or as a consequence of the protein-protein interactions, etc.) making the corresponding monoclonal antibody less specific and less avid. The various methods, ELISA, WB and IHC, seeking the same molecule, they use antibodies that not individualize the same epitope on molecule target, for different state (native or modified) of the inducer molecular antigen at correspondent antibody formation.