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Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab129195 as a replacement.
Our Abpromise guarantee covers the use of ab53269 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 93 kDa).|
Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
Lane 2: KDM1A knockout HAP1 whole cell lysate (40 µg)
Lane 3: HeLa whole cell lysate (40 µg)
Lane 4: Jurkat whole cell lysate (40 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab53269 observed at 92 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab53269 was shown to specifically react with KDM1 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KDM1 knockout samples were used. Wild-type and KDM1 knockout samples were subjected to SDS-PAGE. Ab53269 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 2 µg/ml concentration and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.
ab53269 staining KDM1A in wild-type HAP1 cells (top panel) and KDM1A knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab53269 at 1μg/ml concentration and ab202272 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
ab53269 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"