Anti-NQO1 antibody

• ICC

Lab

Immunocytochemistry - Anti-NQO1 antibody (AB34173)

ab34173 staining NQO1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab34173 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Lab

Western blot - Anti-NQO1 antibody (AB34173)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : NQO1 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HepG2 whole cell lysate (20 μg)
Lane 4 : MCF7 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab34173 observed at 31 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab34173 was shown to specifically react with NQO1 in wild-type HAP1 cells as signal was lost in NQO1 knockout cells. Wild-type and NQO1 knockout samples were subjected to SDS-PAGE. ab34173 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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Western blot - Anti-NQO1 antibody (ab34173)

Predicted band size: 31 kDa

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• WB

Project

Western blot - Anti-NQO1 antibody (AB34173)

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Western blot - Anti-NQO1 antibody (ab34173) at 1 µg/mL

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Human Kidney Whole Tissue Lysate at 20 µg

Secondary

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IRDye 680 Conjugated Goat Anti Rabbit IgG (H&L) at 1/15000 dilution

Predicted band size: 31 kDa

Observed band size: 31 kDa

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• WB

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Western blot - Anti-NQO1 antibody (AB34173)

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Western blot - Anti-NQO1 antibody (ab34173) at 1 µg/mL

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Recombinant human NQO1 protein (<a href='/en-us/products/unavailable/recombinant-human-nqo1-protein-ab59663'>ab59663</a>) at 0.01 µg

Secondary

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Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 31 kDa

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Exposure time: 30s