Immunofluorescence analysis of proNGF expression in human DU145 prostate cancer cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) DU145 cells were incubated with proNGF antibody S-080-100 (1:1000, red) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-sheep-PE antibody (1:100, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.
The sheep anti-proNGF antibodies recognize recombinant proNGF, the prodomain of NGF and but not mouse mature NGF (50 ng in each lane).