1. Characterization of monoclonal antibody 4
Characterization of monoclonal antibody 4.47 for specific p125FAK immunoreactivity.
Characterization of monoclonal antibody 4.47 for specific p125FAK immunoreactivity. A, left, indirect immunofluorescence on BT-474 breast cancer cells demonstrates intense staining at focal adhesions; middle, the FAK −/− knockout fibroblasts demonstrate no focal adhesion staining with 4.47, despite having intact focal adhesions by paxillin staining (right), thus demonstrating the specificity of the antibody. B, Western blot analyses comparing antibodies 4.47 and C20 on lysates of RD rhabdomyosarcoma cells, BT-474 cells, and FAK−/− fibroblasts. An intense Mr 125,000 band is demonstrated in the RD and BT-474 cells, with no reactivity in the FAK knockout fibroblasts. C, immunoprecipitation analyses demonstrating that antibody 4.47 immunoprecipitates a Mr 125,000 protein from a RD cell lysate, detected by both 4.47 and C20. The murine IgG heavy chain (IgGHC) is seen in the 4.47 lane.
William G. Cance et al. Clin Cancer Res 2000;6:
©2000 by American Association for Cancer Research