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Synthetic peptide corresponding to residues near the N-terminus of human alpha 1 Catenin.
A trial size is available to purchase for this antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
Our Abpromise guarantee covers the use of ab51032 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/50000. Detects a band of approximately 100 kDa (predicted molecular weight: 100 kDa).|
|ICC/IF||1/100 - 1/250.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration.|
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.
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.
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 4oC. The secondary antibody was a Goat Anti-Rabbit Alexa Fluor 488 (IgG H&L; ab150077) used at a 1/500 dilution.
Overlay histogram showing MCF7 cells stained with ab51032 (red line). The cells were fixed with 4% PFA (10 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 (ab51032, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was goat anti-rabbit DyLight® 488 (IgG H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/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 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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