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Recombinant fragment: RVFEISPFEP WITRDKVERM HITDMKLPHL PGLEDLGIQA TPLELKAIEV LRRHRTYRWL SAEIEDVKPA KTVNI, corresponding to amino acids 303-378 of Human NDUFA9
Our Abpromise guarantee covers the use of ab55521 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 43 kDa.|
|IHC-P||Use a concentration of 8 µg/ml.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|Flow Cyt||Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, 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)
Lanes 1 - 2: Merged signal (red and green). Green - ab55521 observed at 40 kDa. Red - loading control, ab18251, observed at 52 kDa.
ab55521 was shown to recognize NDUFA9 when NDUFA9 knockout samples were used, along with additional cross-reactive bands. Wild-type and NDUFA9 knockout samples were subjected to SDS-PAGE. ab55521 and ab18251 (loading control to alpha tubulin) were diluted at 1 µg/mL and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemical analysis of formalin fixed and paraffin embedded human small Intestine tissue labeling NDUFA9 with ab55521 at 8ug/mL.
Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 cells labeling NDUFA9 with ab55521 at 10 ug/mL.
Overlay histogram showing HepG2 cells stained with ab55521 (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 (ab55521, 0.5µg/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 IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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