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A synthetic peptide corresponding to residues on the N-terminus of human alpha Actinin
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 ab68194 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000 - 1/10000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).|
|Flow Cyt||1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Alpha Actinin knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: NIH 3T3 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab68194 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab68194 was shown to specifically react with alpha Actinin in wild-type HAP1 cells as signal was lost in alpha Actinin knockout cells. Wild-type and alpha Actinin knockout samples were subjected to SDS-PAGE. ab68194 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
ab68194 staining alpha Actinin in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 30% BSA for 10 minutes at 37°C; antigen retrieval was by heat mediation in EDTA. Samples were incubated with primary antibody (1/100 in PBS) for 16 hours at 4°C. ab80436 EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit was used.
Overlay histogram showing HeLa cells stained with ab68194 (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 (ab68194, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG; H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ab68194 has not yet been referenced specifically in any publications.