Overlay histogram showing Raji cells stained with ab124945 (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 (ab124945, 1/1000 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) (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.
![All lanes : Anti-IRE1 (phospho S724) antibody [EPR5253] (ab124945) at 1/1000 dilutionLane 1 : Untreated 293T cell lysateLane 2 : Calyculin A treated 293T cell lysateLysates/proteins at 10 µg per lane.Secondarystandard HRP labelled goat anti-rabbit at 1/2000 dilutiondeveloped using the ECL technique](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/9910_IRE1-Primary-antibodies-ab124945-1.jpg)
All lanes : Anti-IRE1 (phospho S724) antibody [EPR5253] (ab124945) at 1/1000 dilutionLane 1 : Untreated 293T cell lysateLane 2 : Calyculin A treated 293T cell lysateLysates/proteins at 10 µg per lane.Secondarystandard HRP labelled goat anti-rabbit at 1/2000 dilutiondeveloped using the ECL technique
Equilibrium disassociation constant (KD)Learn more about KD Click here to learn more about KD