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Our Abpromise guarantee covers the use of ab19487 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 at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 91 kDa).|
|ICC||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|IHC (PFA fixed)||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration. PubMed: 23576758|
|Flow Cyt||Use 0.1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Empty (0 µg)
Lane 3: BCL11A (Ctip1) knockout HAP1 whole cell lysate (20 µg)
Lane 4: Raji whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab19487 observed at 91 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab19487 was shown to specifically react with BCL11A (Ctip1) in HAP1 wild type cells. No band was observed when BCL11A (Ctip1) knockout cells were examined. Wild-type and BCL11A (Ctip1) knockout samples were subjected to SDS-PAGE. ab19487 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution 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 1 hour at room temperature before imaging.
The CTIP1 protein band is indicated at 92 kDa. This antibody also identified a 55 kDa protein band in Western blotting, principally in mouse fibroblasts (MEF), but this is unrelated to.
Blocking was performed in 5% milk (in PBS). The primary antibody was diluted in 1% milk (in PBS) and incubated overnight at 4ºC.
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