Anti-alpha 1 Catenin 抗体 [EP1793Y] (ab51032)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Alpha 1 Catenin 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 alpha 1 Catenin in wild-type cells as signal was lost at the expected MW in alpha 1 Catenin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and alpha 1 Catenin 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.

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

ICC/IF image of ab51032 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed in 4% formaldehyde (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the primary antibody (ab51032, 1 µg/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 1 hour. An Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

ICC/IF image of anti-alpha 1 Catenin 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.

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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.