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Our Abpromise guarantee covers the use of ab58668 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent dilution. Detects a band of approximately 35 kDa (predicted molecular weight: 21 kDa).|
|Dot Blot||Use at an assay dependent dilution.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration. PubMed: 20054152|
ab58668 staining ATF3 in A549 cells treated with ionomycin Ca2+ salt (ab120116), by ICC/IF. Increase in ATF3 expression correlates with increased concentration of ionomycin Ca2+ salt, as described in literature.
The cells were incubated at 37°C for 2h in media containing different concentrations of ab120116 (ionomycin Ca2+ salt) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab58668 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
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