Anti-NDUFA9 antibody [20C11B11B11]

• WB

Lab

Western blot - Anti-NDUFA9 antibody [20C11B11B11] (AB14713)

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 was shown to specifically react with NDUFA9 in wild-type HAP1 cells. No band was observed when NDUFA9 knockout HAP1 samples were used. Wild-type and NDUFA9 knockout samples were subjected to SDS-PAGE. ab14713 and ab176560 (loading control to alpha tubulin) were diluted at 1μg/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 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

Predicted band size: 43 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFA9 antibody [20C11B11B11] (AB14713)

ab14713 a 1/100 dilution staining NDUFA9 in Human spinal column tissue by Immunohistochemistry (Formalin/PFA-Fixed paraffin-embedded sections). Antibody was incubated with the sample for 1 hour. Sections were incubated in peroxidase-conjugated rabbit anti-mouse secondary (diluted 1/100 in 4% BSA in PBST) for 1 hour at room temperature. Sections were washed x3 in PBST and peroxidase activity was demonstrated using kit. Antigen retrieval was performed by 1 minute of pressure cooking with 1 mmol EDTA pH 8.0.

• Flow Cyt

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Flow Cytometry - Anti-NDUFA9 antibody [20C11B11B11] (AB14713)

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 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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.

• WB

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Western blot - Anti-NDUFA9 antibody [20C11B11B11] (AB14713)

The band observed at 36 kDa could potentially be a cleaved form of NDUFA9 due to the presence of a 35 amino acid transit peptide.

All lanes:

Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713) at 1 µg/mL

Lane 1:

WI38 (Human lung fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 2:

Human testis tissue lysate - total protein (<a href='/en-us/products/unavailable/human-testis-tissue-lysate-total-protein-ab30257'>ab30257</a>) at 10 µg

Lane 3:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 43 kDa

Observed band size: 36 kDa,58 kDa

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Exposure time: 20min

• WB

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Western blot - Anti-NDUFA9 antibody [20C11B11B11] (AB14713)

All lanes:

Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

Lane 1:

Isolated mitochondria from Human heart at 5 µg

Lane 2:

Isolated mitochondria from Bovine heart at 1 µg

Lane 3:

Isolated mitochondria from Rat heart at 10 µg

Lane 4:

Isolated mitochondria from Mouse Heart at 10 µg

Secondary

All lanes:

Goat anti-Mouse IgG

Predicted band size: 43 kDa

Observed band size: 37 kDa

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