Immunofluorescence analysis of proGDNF expression in C6 (rat glial tumor cell line) cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X-100) SH-SY5Y cells were incubated with ab244211 at 2 μg/mL, (green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue).

Immunofluorescence analysis of proGDNF expression in SH-SY5Y (human neuroblastoma cell line from bone marrow) cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X-100) SH-SY5Y cells were incubated with ab244211 at 2 μg/mL, (green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue).

Fixing and permeabilization of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: ab244211 at 2 μg per ~10⁶ cells for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1/100) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 was used as negative control under same conditions (black dashed).