Immunohistochemistry, western blot and immunoprecipitation analyses of CSF1R.

<p>(A) IHC staining of antibody FER216 labelled the HEK-CSFR1-Fc transfectants expressing CSFR1 but not the negative control HEK-FGFR1-Fc expressing Fc transfectants. The positive anti-Fc mAb control labelled both sets of transfectants. (B) Western blot analyses of CSF1R protein using protein extracts from HEK293 cells transfected with either CSF1R or a closely related protein (FGFR1) as a negative control, normal tissues, cHL tumour samples, and cHL-derived cell lines. Antibody FER216 recognised a band of 100 kDa in tonsil extracts and two bands of 100 and 150 kDa in extract from three cHL biopsies. Comparable results were obtained using the anti-CSFR1 mAb clone 61701. The mAb to tubulin was used as the loading control. (C) IP followed by western blotting demonstrated comparable results using either the FER216 or Clone 61701. Both antibodies were able to immunoprecipitate bands of 100 kDa from tonsil and 100 and 150 kDa from a case of cHL.