Anti-NDUFS3 antibody [17D95]

• Flow Cyt

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Flow Cytometry - Anti-NDUFS3 antibody [17D95] (AB14711)

Overlay histogram showing HepG2 cells stained with ab14711 (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 (ab14711, 1μg/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 IgG2a [ICIGG2A] (ab91361, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• WB

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Western blot - Anti-NDUFS3 antibody [17D95] (AB14711)

The band observed at 26 kDa could potentially be a cleaved form of NDUFS3 due to the presence of a 36 amino acid transit peptide.

All lanes:

Western blot - Anti-NDUFS3 antibody [17D95] (ab14711) at 1 µg/mL

Lane 1:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 2:

Human heart tissue lysate - total protein (<a href='/en-us/products/unavailable/human-heart-tissue-lysate-total-protein-ab29431'>ab29431</a>) at 10 µg

Lane 3:

Heart (Mouse) Tissue Lysate at 10 µg

Lane 4:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 30 kDa

Observed band size: 26 kDa,57 kDa

true

Exposure time: 1min

• WB

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Western blot - Anti-NDUFS3 antibody [17D95] (AB14711)

Extra bands in the mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.

All lanes:

Western blot - Anti-NDUFS3 antibody [17D95] (ab14711)

Lane 1:

Isolated mitochondria from Human heart at 5 µg

Lane 2:

Isolated mitochondria from Bovine heart at 4 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: 30 kDa

Observed band size: 26 kDa

false

• WB

AbReview51457****

Western blot - Anti-NDUFS3 antibody [17D95] (AB14711)

All lanes:

Western blot - Anti-NDUFS3 antibody [17D95] (ab14711) at 1/1000 dilution

Lanes 1 - 2:

Whole cell lysates from NDUFS3 KO cells. at 50 µg

Lanes 3 - 4:

Whole cell lysates from NDUFS3 WT cells. at 50 µg

Lanes 5 - 6:

Isolated mitochondria from NDUFS3 KO cells. at 50 µg

Lanes 7 - 8:

Isolated mitochondria from NDUFS3 WT cells. at 50 µg

Secondary

All lanes:

Goat anti-mouse polyclonal HRP conjugate at 1/5000 dilution

Predicted band size: 30 kDa

Observed band size: 22 kDa,40 kDa

true

Exposure time: 1min

Image is courtesy of an anonymous AbReview