The importance of characterization of FITC-labeled antibodies used in tissue cross-reactivity studies.

Takai, H., Kato, A., Nakamura, T., Tachibana, T., Sakurai, T., Nanami, M., Suzuki, M.
Journal   Acta Histochem.
Species  
Analytes Measured   IL-6R
Matrix Tested   Peripheral blood mononuclear cell (PBMC) culture supernatants
Year   2010
Volume  
Page Numbers  
Application   Cytokines and Chemokines
Abstract
Fluorescein isothiocyanate (FITC)-labeled antibodies are widely used as primary antibodies in the tissue cross-reactivity (TCR) studies for the development of therapeutic antibodies. However, the effects of FITC-labeling on the characteristics of an antibody are poorly understood. The present study was performed to examine the effect of FITC-labeling on the binding affinity and immunohistochemical staining profile of an antibody, using several antibodies with different FITC-labeling indices. The results showed that the FITC-labeling index in antibody was negatively correlated with the binding affinity for its target antigen. Immunohistochemically, an antibody with a higher labeling index had a tendency to be more sensitive, but was also more likely to yield non-specific staining. Based on these findings, we recommend that a FITC-labeled antibody used as a primary antibody in a TCR study should be carefully selected from several differently labeled antibodies to minimize the decrease in the binding affinity and achieve the appropriate sensitivity and interpretation of the immunohistochemistry.

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