Anti-CTNNA1 antibody [EP1793Y] - Low endotoxin, Azide free (ab226010)

This WB data was generated using the same anti-CTNNA1 antibody clone [EP1793Y] in a different buffer formulation (cat# ab51032).

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: CTNNA1 HAP1 whole cell lysate (20 µg) 
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab51032 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab51032 was shown to recognize CTNNA1 in wild-type cells as signal was lost at the expected MW in CTNNA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CTNNA1 knockout samples were subjected to SDS-PAGE. Ab51032 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

ICC/IF image of anti-CTNNA1 antibody [EP1793Y] (ab51032) stained mouse ES cells. The cells were fixed in 1:1 methanol/acetone, permeabilized using 0.1% Triton X, and blocked with 1% serum in PBS for 30 minutes. The cells were then incubated with ab51032 at a 1/100 dilution for 16 hours at 4^oC. The secondary antibody was a Goat Anti-Rabbit Alexa Fluor^® 488 (IgG H&L;  ab150077) used at a 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51032).

Paraffin-embedded human breast carcinoma tissue stained for CTNNA1 with ab51032 at a 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51032).

Overlay histogram showing MCF7 (Human breast adenocarcinoma cell line) cells stained with ab51032 (red line). The cells were fixed with 4% PFA (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 primary antibody (ab51032, 1/100 dilution) for 30 minutes at 22ºC. The secondary antibody used was goat anti-rabbit DyLight^® 488 (IgG H+L) ab96899 at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1 µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51032).

This ICC/IF data was generated using the same anti-CTNNA1 antibody clone [EP1793Y] in a different buffer formulation (cat# ab51032).

ICC/IF image of ab51032 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51032, 1µg/ml) overnight at +4°C. The secondary antibody (green) was anti-rabbit Alexa Fluor® 488 (IgG H+L; ab150077) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.