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Immunofluorescence analysis of NQO1 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with NQO1 (A180) Mouse Monoclonal Antibody (39-3700) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Dog, Human, Non-human primate, Rat|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant human NQO1 protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||2-4 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
NQO1 is a member of the NAD(P)H dehydrogenase(quinone) family and a cytoplasmic 2-electron reductase. This FAD-binding protein forms homodimers and reduces quinones to hydroquinones. This protein's enzymatic activity prevents the one electron reduction of quinones that results in the production of radical species. Altered expression of the protein has been seen in many tumors and is also associated with Alzheimer's disease (AD).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cross-regulations among NRFs and KEAP1 and effects of their silencing on arsenic-induced antioxidant response and cytotoxicity in human keratinocytes.
39-3700 was used in western blot to study the regulatory interactions of NRF2, NRF, and KEAP and their role in the anti-oxidant responses of human keratinocytes to arsenic toxicity.
|Zhao R,Hou Y,Zhang Q,Woods CG,Xue P,Fu J,Yarborough K,Guan D,Andersen ME,Pi J||Environmental health perspectives (120:583)||2012|
Prolonged inorganic arsenite exposure suppresses insulin-stimulated AKT S473 phosphorylation and glucose uptake in 3T3-L1 adipocytes: involvement of the adaptive antioxidant response.
39-3700 was used in western blot to report that arsenic exposure activates the adaptive oxidative stress response and impairs insulin-stimulated ROS signaling, and thus causes insulin resistance in adipocytes.
|Xue P,Hou Y,Zhang Q,Woods CG,Yarborough K,Liu H,Sun G,Andersen ME,Pi J||Biochemical and biophysical research communications (407:360)||2011|
|Human||Not Cited||Interaction of human NAD(P)H:quinone oxidoreductase 1 (NQO1) with the tumor suppressor protein p53 in cells and cell-free systems.||Anwar A,Dehn D,Siegel D,Kepa JK,Tang LJ,Pietenpol JA,Ross D||The Journal of biological chemistry (278:10368)||2003|
||Interaction of human NAD(P)H:quinone oxidoreductase 1 (NQO1) with the tumor suppressor protein p53 in cells and cell-free systems.||Anwar A,Dehn D,Siegel D,Kepa JK,Tang LJ,Pietenpol JA,Ross D||The Journal of biological chemistry (278:10368)||2003|
Azoreductase; DHQU; DIA4; Diaphorase (NADH/NADPH); diaphorase (NADH/NADPH) (cytochrome b-5 reductase); diaphorase-4; dioxin-inducible 1; DT-diaphorase; DTD; Menadione reductase; NAD(P)H dehydrogenase [quinone] 1; NAD(P)H dehydrogenase, quinone 1; NAD(P)H:menadione oxidoreductase; NAD(P)H:menadione oxidoreductase 1; NAD(P)H:Quinone acceptor oxidoreductase type 1; NAD(P)H:quinone oxidoreductase 1; NAD(P)H:quinone oxireductase; NMOR1; NMORI; NQ01; Phylloquinone reductase; QR1; Quinone reductase 1
DHQU; DIA4; DTD; NMOR1; NMORI; NQO1; QR1