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Active NADH Dehydrogenase, Quinone 1 (NQO1)

DHQU; DIA4; DTD; NMOR1; NMORI; QR1; Azoreductase; DT-diaphorase; Menadione reductase; Phylloquinone reductase; Quinone reductase 1; NAD(P)H:quinone oxidoreductase 1

ACTIVITY TEST

NAD(P)H:quinone acceptor oxidoreductase 1 (NQO1), also known as DT-diaphorase, is a widely-distributed FAD-dependent flavoprotein that promotes 2-electron reductions of quinones, quinoneimines, nitroaromatics, and azo dyes. As a result it prevents the one electron reduction of quinones that results in the production of radical species. NQO1 is a highly-inducible enzyme that is regulated by the Keap1/Nrf2/ARE pathway. The increase and decrease of NQO1 levels are associated with decreased and increased susceptibilities to oxidative stress, respectively. Thus, NQO1 is a marker cytoprotective enzyme in oxidative stress. Independently of its catalytic function, NQO1 plays a role in regulating the proteosomal degradation of p53, p73a, and p33. NQO1 physically interacts with p53 and p73 in an NADH-dependent manner and protects them from 20S proteasomal degradation in a ubiquitin independent pathway. The activity assay of recombinant human NQO1 was measured by its ability to oxidize the substrate resazurin to resorufin. The rhNQO1 was diluted to 100 ug/ml in the assay buffer 50 mM HEPES, 0.2 M NaCl, 5 µM FAD, 0.05% Tween® 20, pH 7.5. 50 ul 100 ug/ml rhNQO1 was added into the microplate and start the reaction by adding 50 µl substrate mixture of 400 uM beta-NADH and 20 uM resazurin which was diluted in assay buffer. Read at excitation and emission wavelengths of 540 nm and 585 nm (top read), respectively, in kinetic mode for 5 minutes. The specific activity of recombinant human NQO1 is >18 pmol/min/µg.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Hypertension Transcription factor Nrf2 protects renal dopamine D1 receptor function during oxidative stress. Pubmed: 23876469
Free Radical Research Notoginsenoside R1-mediated neuroprotection involves estrogen receptor-dependent crosstalk between Akt and ERK1/2 pathways: A novel mechanism of Nrf2/ARE signaling activation Informahealthcare: Source
physiological reports Antioxidant resveratrol restores renal sodium transport regulation in SHR PubMed: 26603454
Biol Chem Genetic association of NAD (P) H Quinone Oxidoreductase (NQO1* 2) polymorphism with NQO1 levels and risk of diabetic nephropathy Pubmed:27078674
Experimental Parasitology Sodium arsenite augments sensitivity of Echinococcus granulosus protoscoleces to albendazole Pubmed: 30790573
International Journal of Nanomedicine Gold Nanoparticles Perturb Drug-Metabolizing Enzymes and Antioxidants in the Livers of Male Rats: Potential Impact on Drug Interactions Pubmed: 32764932
Exp Parasitol In vitro protoscolicidal effects of lithocholic acid on protoscoleces of Echinococcus granulosus and its mechanism Pubmed:35594934
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