IF staining of U251 cells using 033127. Cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with antibody 033127 (1:25 dilution; 1 h at 37°C). Alexa Fluor® 488-conjugated donkey anti-rabbit secondary antibody (green) was used (1:400, 50 min at 37°C). Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (7units/ml, 1h at 37°C). CACNA2D2 immunoreactivity is localized significantly to cytoplasm.