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Other Immunogen Type corresponding to Human DNA PKcs aa 1-2713. Immunogen is purified from HeLa cells.
Database link: P78527
Our Abpromise guarantee covers the use of ab44815 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Inhibition Assay||Use at an assay dependent concentration.
Inhibits (50%) DNA-dependent phosphorylation.
|WB||1/200. Predicted molecular weight: 469 kDa.|
Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: DNA PKcs knockout HAP1 cell lysate (40 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: SHSY5Y cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab44815 observed at 460 kDa. Red - loading control, ab18251, observed at 52 kDa.
ab44815 was shown to specifically react with DNA PKcs when DNA PKcs knockout samples were used. Wild-type and DNA PKcs knockout samples were subjected to SDS-PAGE. Ab44815 and ab18251 (loading control to alpha tubulin) were diluted at 1/200 and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
ab44815 staining DNA PKcs in wild-type HAP1 cells (top panel) and PRKDC 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 ab44815 at 1/100 dilution and ab202272 at 1/250 dilution (shown in pseudocolour 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.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab44815 has not yet been referenced specifically in any publications.