Considerations for establishing the validity of immunocytological studies

Immunocytology has wide spread applications for localizing tissue antigens, as evidenced by the recent exploitation of this technique in biological studies. Documenting the immunological specificity of the staining reaction is one of the most important technical considerations in validating the accrued data in immunocytological studies. The purpose of this report is to discuss and emphasize the need for conducting physiological studies in addition to the traditional immunological method and specificity controls. The ability of antibodies to bind molecules other than those molecules used as the immunizing material is a well documented fact. Hypothetically, preabsorption of the primary antibody with its specific antigen, could reduce subsequent binding of this antibody to a cross reactive tissue antigen, thus providing false confirmation of staining validity. The results of our experience with a cross reacting system in addition to other previously reported examples are discussed.