Recombinant Human NQO-1 Protein, CF

R&D Systems | Catalog # 7567-DH

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human NQO-1 Protein (7567-DH)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P15559

Applications

Enzyme Activity
View all NQO-1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human NQO-1 protein
Met1-Lys274, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met1

Predicted Molecular Mass

32 kDa

SDS-PAGE

29-31 kDa, reducing conditions

Activity

Measured by its ability to oxidize Resazurin.
The specific activity is >15,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7567-DH
Formulation Supplied as a 0.2 μm filtered solution in NaH2PO4 and NaCl.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: NQO-1

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 (1, 2). 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 (2, 3). 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 (2). NQO1 physically interacts with p53 and p73 in an NADH-dependent manner and protects them from 20S proteasomal degradation in a ubiquitin independent pathway (4).

References

  1. Gong, X. et al. (2008) Vitamin Horm. 78:85.
  2. Dinkova-Kostova, A. T., and Talalay, P. (2010) Arch. Biochem. Biophys. 501:116.
  3. Hayes, J. D. and McMahon, M. (2009) Trends Biochem. Sci. 34:176.
  4. Asher, G. et al. (2005) Genes Dev. 19:316.

Long Name

NAD(P)H Dehydrogenase, Quinone 1

Alternate Names

Azoreductase, DHQU, DIA4, Diaphorase-4, Dioxin-inducible 1, DTD, Menadione Reductase, NMOR1, NQO1, QR1, Quinone Reductase 1

Entrez Gene IDs

1728 (Human)

Gene Symbol

NQO1

UniProt

P15559 (Human)

Additional NQO-1 Products

Product Documents for Recombinant Human NQO-1 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human NQO-1 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd. Tween is a registered trademark of ICI Americas.

For research use only

Related Research Areas

p53 Pathway
Redox Enzymes

Citations for Recombinant Human NQO-1 Protein, CF

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Protocols

View specific protocols for Recombinant Human NQO-1 Protein, CF (7567-DH):

Materials
  • Assay Buffer: 50 mM HEPES, 0.2 M NaCl, 5 µM FAD, 0.05% Tween® 20, pH 7.5
  • Recombinant Human NQO-1 (rhNQO-1) (Catalog # 7567-DH)
  • beta -Nicotinamide Adenine Dinucleotide, reduced (NADH) (Sigma, Catalog # N8129), 20 mM stock in  0.1 M Sodium Borate, pH 9.0
  • Resazurin (Catalog # AR002) (MW = 251.17 Da)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhNQO-1 to 0.0075 µg/mL in Assay Buffer.
  2. Dilute NADH to 800 µM in Assay Buffer.
  3. Dilute Resazurin to 40 µM in Assay Buffer.
  4. Combine equivalent volumes of diluted NADH and Resazurin to make the Substrate Mixture. Mix well and use immediately.
  5. Load 50 µL of 0.0075 µg/mL rhNQO-1 into a plate, and start the reaction by adding 50 µL of the Substrate Mixture. For a Substrate Blank, load 50 µL of Assay Buffer and 50 µL of Substrate Mixture.
  6. Read at excitation and emission wavelengths of 540 nm and 585 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard Resorufin (Sigma, Catalog # R3257).

Per Well:

  • rhNQO-1: 0.375 ng
  • NADH: 200 µM
  • Resazurin: 10 µM

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