ab174437 staining L2/HNK-1 Carbohydrate epitope in 4 dpf zebrafish retina sections by IHC-Fr. The tissue was fixed with paraformaldehyde and an antigen retrieval step was performed with sodium citrate pH 6. Blocking of the sample was done with 5% BSA in PBS containing 01% Tween 20 and 0.5% Triton X, for 60 minutes at 23°C, followed by staining with ab174437 at 1/500 in blocking solution for 16h at 4°C. An alexa 647 conjugated goat anti-mouse polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei are stained in blue with DAPI. L2/HNK-1 Carbohydrate epitope stains the inner nuclear layer and inner plexiform layer (red). See Abreview
Immunocytochemistry with SH-SY5Y cells stained with ab174437 in red and DAPI in blue, as a counter stain. Fixation: Paraformaldehyde fixed (4%, 20 min) Permeabilization: Triton X-100 permeabilized (0.1%, 10 min) Blocking: 10% Goat Serum/PBS (1 hour) Primary: ab174437 5 µg/mL (10% Goat Serum/PBS, 2hr at room temperature or overnight at 4°C). Secondary: Alexa 594 goat anti-mouse IgG1 used at a 1/1000 dilution in 10% Goat Serum/PBS for 1hr at RT. Washing: 3x 1% Goat Serum/PBS, 10 minutes/wash DAPI: 2 µM in 1% Goat Serum/PBS, 10 minutes.