beta Actin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to beta Actin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab125248.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.Band: 42kDa; beta Actin
![Overlay histogram showing HeLa cells stained with ab125248 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab125248, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse Alexa Fluor® 488 (IgG; H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 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.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/13656_ab125248-2-ab125248FC.jpg)
Overlay histogram showing HeLa cells stained with ab125248 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab125248, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse Alexa Fluor® 488 (IgG; H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 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.
![All lanes : Anti-beta Actin antibody [BA3R] (ab125248) at 1/1000 dilutionLane 1 : Human tissue lysateLane 2 : Mouse tissue lysateLane 3 : Rat tissue lysateLane 4 : Rabbit tissue lysateLysates/proteins at 20 µg per lane.developed using the ECL technique](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/13657_beta-Actin-Primary-antibodies-ab125248-1.jpg)
All lanes : Anti-beta Actin antibody [BA3R] (ab125248) at 1/1000 dilutionLane 1 : Human tissue lysateLane 2 : Mouse tissue lysateLane 3 : Rat tissue lysateLane 4 : Rabbit tissue lysateLysates/proteins at 20 µg per lane.developed using the ECL technique