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Our Abpromise guarantee covers the use of ab19304 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Detects a band of approximately 370 kDa.|
|IP||Use at an assay dependent concentration.|
|ICC/IF||1/50 - 1/1000.|
Immunofluorescent analysis of ATM phosphorylated on pSer1981 (green) in HeLa cells left untreated (left panels) or treated with 10uM Camptothecin for 4 hours (right panels). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a phospho-ATM (pSer1981) monoclonal antibody (ab19304), at a dilution of 1:1000 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (top panels). A merged image with nuclei staining (blue) using Hoechst 33342 dye is shown on bottom panels. Images were taken at 20X magnification.
Immunofluorescent analysis of Phospho-ATM pSer1981 in 293 untreated cells (left panel) or stimulated cells with 100 mJ UV (right panel) . Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-ATM pSer1981 Monclonal Antibody (10H11)(ab19304) at a dilution of 1:50 and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.
By Western blot this antibody detects a ~370 kDa protein representing the phospho-ATM kinase in crude lysates from gamma irradiated HeLa cells. In immunofluorescence procedures, ab19304recognizes the phospho-ATM kinase in irradiated human and mouse fibroblasts.
ab19304 has not yet been referenced specifically in any publications.
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