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6xHis-tagged fusion protein, corresponding to amino acids 105-291 of Human IKB alpha.
Our Abpromise guarantee covers the use of ab12134 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 35.6 kDa).|
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: IKB alpha knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab12134 observed at 38 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab12134 was shown to specifically react with IKB alpha in wild type cells as signal was lost in IKB alpha knockout cells. Wild-type and IKB alpha knockout samples were subjected to SDS-PAGE. Ab12134 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml 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.
ab12134 at 1/250 staining human adrenal tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was blocked in serum before incubation with the primary antibody for 30 minutes at 22°C. An HRP conjugated goat anti-mouse antibody was used as the secondary.
ab12134 detects both phosphorylated and non-phosphorylated forms of IKB alpha by Western blot. Jurkat cells (1E7) were treated for indicated time periods with 10 nm/ml PMA, 1