We observed intense intracellular immunofluorescence of rat lung fibroblasts stained with hybridoma culture supernatant containing monoclonal antibodies to human plasma fibronectin, but no pericellular matrix staining. Immunoprecipitation and absorption experiments revealed that this intracellular staining by hybridoma-conditioned medium was due to binding of fibronectin-antifibronectin immune complexes via the fibronectin to intracellular procollagen. The anomalous staining patterns we encountered were not revealed by the usual controls for immunohistochemical specificity, and also occurred in rat tissue sections. This general phenomena--binding of serum antigens present in hybridoma medium to cellular components--could in principle result in artifactual staining with monoclonal antibodies to other serum components, so investigators using monoclonal antibodies should be aware of this new artifact. Our results also demonstrate that fibronectin binds specifically to native procollagen. Monoclonal antibodies may be useful for studying fibronectin-procollagen and other macromolecular interactions.
Specific binding of fibronectin--antifibronectin immune complexes to procollagen: a new pitfall in immunostaining.
J A McDonald, D G Kelley; Specific binding of fibronectin--antifibronectin immune complexes to procollagen: a new pitfall in immunostaining.. J Cell Biol 1 March 1984; 98 (3): 1042–1047. doi: https://doi.org/10.1083/jcb.98.3.1042
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