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.
![All lanes : Anti-alpha Actinin antibody [EP2527Y] (ab68194) at 1/10000 dilutionLane 1 : NIH 3T3 lysate Lane 2 : HeLa lysateLane 3 : C6 lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti-rabbit at 1/2000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/5603_ab68194_wb1.jpg)
All lanes : Anti-alpha Actinin antibody [EP2527Y] (ab68194) at 1/10000 dilutionLane 1 : NIH 3T3 lysate Lane 2 : HeLa lysateLane 3 : C6 lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti-rabbit at 1/2000 dilution
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.See Abreview