Overlay histogram showing HeLa cells stained with ab133448 (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 (ab133448, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody [EPR5683] (ab133448) at 1/1000 dilutionLane 1 : 293 cell lysateLane 2 : Lysate of 293 cells treated with lambda phosphataseLysates/proteins at 10 µg per lane.SecondaryGoat anti-rabbit HRP conjugated antibody at 1/2000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/6584_AMPK-alpha-1-Primary-antibodies-ab133448-1.jpg)
All lanes : Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody [EPR5683] (ab133448) at 1/1000 dilutionLane 1 : 293 cell lysateLane 2 : Lysate of 293 cells treated with lambda phosphataseLysates/proteins at 10 µg per lane.SecondaryGoat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Equilibrium disassociation constant (KD)Learn more about KD Click here to learn more about KD