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Tissue, cells or virus corresponding to Cow NDUFA9.
This monoclonal antibody to NDUFA9 has been knockout validated in Western blot. The expected band for NDUFA9 was observed in wild type cells and the band was not seen in knockout cells.
Our Abpromise guarantee covers the use of ab14713 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 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 40 kDa).|
|Flow Cyt||Use 1-2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: NDUFA9 knockout HAP1 cell lysate (20 µg)
Lane 3: WI38 cell lysate (20 µg)
Lane 4: NIH3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab14713 observed at 40 kDa. Red - loading control, ab176560, observed at 52 kDa.
ab14713 detected the expected band for NDUFA9 in wild-type HAP1 cells and the band was not seen in NDUFA9 knockout HAP1 cells. Wild-type and NDUFA9 knockout samples were subjected to SDS-PAGE. ab14713 and ab176560 (loading control to alpha tubulin) were diluted at 1 μg/mL and 1/10 000 respectively and incubated overnight at 4°C. 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.
Overlay histogram showing HepG2 cells stained with ab14713 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab14713, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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