Rabbit anti calnexin antibody

Cells were fixed in 4% paraformaldehyde and permeabilized in 0.5% phosphate-buffered saline (PBS)/Triton X-100 (PBST). The cells were then blocked with 5% goat serum to decrease non-specific binding for 30 min at room temperature and were incubated with mouse anti-Par-4 antibody (1:100; Abcam, USA), rabbit anti-calnexin antibody (1:150; Abcam), mouse anti-GRP78 antibody (1:100; Abcam), and rabbit anti-calnexin antibody (1:150; Abcam) at 4°C for overnight in a wet box. They were then incubated with the secondary antibodies rhodamine (TRITC) goat anti-mouse IgG (1:100; Jackson, USA) and FITC AffiniPure goat anti-rabbit IgG (1:100; Jackson) for 1 h at room temperature in the dark. After staining with 4′,6-diamidino-2-phenylindole for 40 s, the cells were observed and photographed using a confocal laser scanning microscope.