Rabbit anti nqo1

Manufactured by Santa Cruz Biotechnology

Rabbit anti-NQO1 is a primary antibody that specifically targets the NQO1 (NAD(P)H Quinone Dehydrogenase 1) protein. NQO1 is an enzyme that plays a role in cellular detoxification processes. This antibody can be used to detect and study the expression and distribution of NQO1 in various biological samples.

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Anti-NQO1 (40 citations) Rabbit anti-TM (2 citations)

3 protocols using rabbit anti nqo1

Western Blotting for Protein Expression

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For western blotting, 20–40 μg of total protein from each sample was subjected to SDS-PAGE under reducing conditions. Proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The membranes were blocked for 1 h at room temperature and then incubated overnight at 4 °C with primary antibodies. Primary antibodies used were rabbit anti-COX-2 (Santa Cruz), rabbit anti-iNOS (Santa Cruz), rabbit anti-acetyl-p65 (Cell Signalling), rabbit anti-Total-p65 (Cell Signalling), rabbit anti-MAP2(Cell Signalling), rabbit anti-Nrf2 (Santa Cruz), rabbit anti-HO1 (Santa Cruz), rabbit anti-NQO1 (Santa Cruz), rabbit anti-SIRT1 (Santa Cruz) and rabbit anti-actin (Sigma). Primary antibodies were diluted in Tris-buffered saline (TBS), containing 0.1% Tween 20 (TBS-T) and 1 or 5% BSA. Membranes were incubated with the primary antibody overnight at 4 °C. After extensive washing (three times for 15 min each in TBS-T), proteins were detected by incubation with Alexa Fluor 680 goat anti-rabbit secondary antibody (1:10,000; Life Technologies) at room temperature for 1 h. Detection was done using a LICOR Odyssey Imager. All western blot experiments were carried out at least three times.

Velagapudi R., El-Bakoush A, & Olajide O.A. (2018). Activation of Nrf2 Pathway Contributes to Neuroprotection by the Dietary Flavonoid Tiliroside. Molecular Neurobiology, 55(10), 8103-8123.

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Western Blotting for Protein Expression

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For Western blotting, 20-40 μg of total protein from each sample was subjected to SDS-PAGE under reducing conditions. Proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The membranes were blocked for 1 h at room temperature and then incubated overnight at 4°C with primary antibodies. Primary antibodies used were rabbit anti-COX-2 (Santa Cruz), rabbit anti-iNOS (Santa Cruz), rabbit anti-phospho-IκBα (Santa Cruz), rabbit antiphospho-p65 (Cell Signalling), rabbit anti-p65 (Cell Signalling) rabbit anti-acetyl-p65 (Cell Signalling), rabbit anti-Nrf2 (Santa Cruz), rabbit anti-HO1 (Santa Cruz), rabbit anti-NQO1 (Santa Cruz), and rabbit anti-actin (Sigma). Primary antibodies were diluted in Tris-buffered saline (TBS), containing 0.1% Tween 20 (TBS-T) and 1 or 5% BSA. Membranes were incubated with the primary antibody overnight at 4°C. After extensive washing (three times for 15 min each in TBS-T), proteins were detected by incubation with Alexa Fluor 680 goat anti-rabbit secondary antibody (1:10000; Life Technologies) at room temperature for 1 h. Detection was done using a LICOR Odyssey Imager. All Western blot experiments were carried out at least three times.

Velagapudi R., Kumar A., Bhatia H.S., El-Bakoush A., Lepiarz I., Fiebich B.L, & Olajide O.A. (2017). Inhibition of neuroinflammation by thymoquinone requires activation of Nrf2/ARE signalling. International immunopharmacology, 48.

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Protein Quantification and Western Blot Analysis

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HUVECs were lysed in RIPA buffer, containing proteinase inhibitors and total protein quantified using BCA protein assay kit (Pierce Biotechnology). Proteins (30 μg/well) were denatured and separated by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF) membranes. Blots were blocked for 1 hour in 5% non-fat milk/Tris-Buffered Saline Tween-20 (TBS-T), probed with primary rabbit anti-phospho-protein kinase B (Akt), rabbit anti-Akt (Santa Cruz Biotechnology), rabbit polyclonal anti-phospho-eNOS (Cell Signaling), rabbit anti-eNOS, rabbit anti-HO-1 (Stressgen), rabbit anti-Nrf2, rabbit anti-NQO-1 (Santa Cruz Biotechnology) or mouse anti-β-actin (Sigma), overnight at 4ºC followed by washing and incubation with the corresponding horse radish peroxidase-labelled secondary antibody for 1 hour at room temperature. After washing with TBS-T, membranes were incubated with ECL reagent and visualized (Amersham Pharmacia Biotech).
Densitometry analysis using ImageJ (version 1.32j, NIH, http://rsb.info.nih/ij/) was used to quantify protein signal.

Mahmoud A.M., Wilkinson F.L., Jones A.M., Wilkinson J.A., Romero M., Duarte J, & Alexander M.Y. (2017). A novel role for small molecule glycomimetics in the protection against lipid-induced endothelial dysfunction: Involvement of Akt/eNOS and Nrf2/ARE signaling. Biochimica et biophysica acta. General subjects, 1861(1 Pt A).

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