TrkC expression (red) in 50B11 mouse x rat hybrid cell line analysed by Flow Cytometry. Fixation: 0 ,2 and 4% formaldehyde in PBS, pH 7.2-7.6 on ice (10 minutes). Blocking: 100 µg/mL normal sheep IgG in PBS (30 minutes) on ice. Primary antibody: TrkC clone BS337 (10 µg/mL) for 60 minutes on ice. Secondary antibody: Goat anti-rabbit PE (1:100 dilution), 30 min in dark on ice. Non-specific control IgG clone X63 (black) was used as negative control under same conditions. Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols.
Membrane TrkC expression (green) in human HeLa cells, analyzed by Immunocytochemistry. Fixed (4% formaldehyde) and blocked (10% normal horse serum) HeLa cells were incubated with TrkC antibody M-1837-100 (2 µg/mL, green) for 1 hour. Cells were then permeabilized, blocked (10% horse serum, 0.1% Triton X-100) and incubated with rabbit NF-H antibody R-1389-50 (1:500, red) for 1 hour. Primary antibody binding was visualized with secondary donkey anti-mouse-CF488A and donkey anti-rabbit-CF568 antibodies (2 µg/mL, 1 hour incubation). Cell nuclei were stained with DAPI (blue). Negative control staining was performed under identical conditions with isotype control mouse antibody and rabbit whole serum from an non-immunized animal. Confocal imaging settings were the same for antibody and control staining. Magnification: 60x. Specific staining is observed for TrkC in the cell membrane, while the NF-H antibody reveals intracellular filaments.
3D confocal imaging of membrane TrkC expression (green) in human HeLa cell. TrkC-IR is concentrated in the elongated protrusion away from the cell body, while intracellular NF-H expression (red) is observed throughout the cell. Blue: DAPI nuclear stain.