ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionOccurs in almost all aerobically respiring organisms and serves to protect cells from the toxic effects of hydrogen peroxide. Promotes growth of cells including T-cells, B-cells, myeloid leukemia cells, melanoma cells, mastocytoma cells and normal and transformed fibroblast cells.
Involvement in diseaseDefects in CAT are the cause of acatalasia (ACATLAS) [MIM:115500]; also known as acatalasemia. This disease is characterized by absence of catalase activity in red cells and is often associated with ulcerating oral lesions.
Sequence similaritiesBelongs to the catalase family.
Post-translational modificationsThe N-terminus is blocked.
Western blot - Anti-Catalase antibody [1B6] (ab125688)
Predicted band size : 60 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: Empty Lane 3: Catalase knockout HAP1 cell lysate (20 µg) Lane 4: Empty Lanes 1 - 4: Merged signal (red and green). Green - ab125688 observed at 60 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab125688 was shown to specifically react with Catalase when Catalase knockout samples were used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab125688 and ab181602 (loading control to GAPDH) were diluted 1/200 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Overlay histogram showing HeLa cells stained with ab125688 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab125688, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.