HEK 293 cells were grown on poly-L-lysine-coated coverslips overnight. Cells were then transiently transfected with GPR56. After 24 h, cells were fixed with 4% paraformaldehyde and 0.2% picric acid in phosphate buffer (pH 6.9) for 30 min at room temperature and washed several times with phosphate buffer. Specimens were permeabilized with ice-cold methanol and then incubated with Anti-GPR56 Rabbit Polyclonal Antibody (Product # 720373 at a concentration of 2 µg/mL). Cells were then incubated with Alexa488-conjugated secondary antibody for 2 h at room temperature, mounted and examined using a laser scanning confocal microscope. Note, strong staining of transfected cells and no staining of adjacent non-transfected cells.

Sections of human small intestine were dewaxed, microwaved in citric acid and incubated with Anti-GPR56 Rabbit Polyclonal Antibody (Product # 720373 at a concentration of 2 µg/mL). Sections were then sequentially treated with biotinylated anti-rabbit IgG and AB solution. Sections were then developed in AEC and lightly counterstained with hematoxylin. Inset, for adsorption controls the antibody was incubated with 10 µg/mL of the peptide used for immunizations (+ Peptide). Note, staining of neuroendocrine cells is completely neutralized by preincubation of the antibody with its immunizing peptide.