Anti-Calnexin 抗体 [6F12BE10] (ab112995)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Calnexin knockout HAP1 cell lysate (20 µg)
Lane 3: THP1 cell lysate (20 µg)
Lane 4: Raw264.7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab112995 observed at 80 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab112995 was shown to specifically react with Calnexin. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab112995 at a concentration of 1 µg/mL and ab181602 (loading control to GAPDH) diluted to 1/1000 were incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

ab112995 staining Calnexin (shown in green) in wild-type HAP1 cells (top panel) and CANX knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 min and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab112995 at 0.5 μg/ml and ab202272 at 1/250 dilution (alpha tubulin shown in red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with a goat secondary antibody to Mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab112995 at 5µg/ml staining Calnexin - ER membrane marker in Human fibroblasts cells by Immunocytochemistry (4% paraformaldehyde fixed and 0.1% Triton X-100 permeabilized) followed by Alexa Fluor^® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h (green).