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AMYTLPRQATPGVPAQ, corresponding to amino acids 832-847 of Human LSD1
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 recognize KDM1 when KDM1 knockout samples were used, along with additional cross-reactive bands. 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 dilution and 1/10000 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 1/10000 dilution for 1 hour 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.
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