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D1315

Sigma-Aldrich

DT Diaphorase (NQO1) human

lyophilized powder, recombinant, expressed in E. coli, ≥90% (SDS-PAGE)

Synonym(s):

DTD, NQO1, Quinone reductase

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1 MG
€343.00

About This Item

Enzyme Commission number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

€343.00


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Request a Bulk Order

recombinant

expressed in E. coli

Quality Level

Assay

≥90% (SDS-PAGE)

form

lyophilized powder

specific activity

≥100 units/mg protein

mol wt

monomer 31000

UniProt accession no.

storage temp.

2-8°C

Gene Information

human ... NQO1(1728)

Application

Human DT diaphorase has been used in a study to assess the development of novel quinone phosphorodiamidate prodrugs. Human DT diaphorase has also been used to investigate its crystal structure for the development of a model for its interaction with the cytotoxic prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954).

Biochem/physiol Actions

DT-diaphorase, also referred to as NAD(P)H:(quinone-acceptor) oxidoreductase, is involved in the reductive activation process of several cytotoxic antitumor quinones and nitrobenzenes. It catalyzes the two-electron reduction of quinones and quinonoid compounds to hydroquinones, using either NADH or NADPH as the electron donor. The flavoenzyme contains one mole of FAD per mole of enzyme.[1]
NQO1 is a cytosolic homodimeric FAD-dependent enzyme that catalyses the reduction of a broad range of cytotoxic quinones resulting in protection from cellular oxidative stress. Oxidative stress may also enhance NQO1-mediated protection of p53 and p73 against proteasomal degredation. The highly Inducible expression of NQO1 is controlled by the Nrf2-Keap1/ARE pathway and appears to be affected by changes in susceptibility to oxidative stress. During Oxygen/glucose deprivation, NQO1 appears to be involved in AMPK-induced cancer cell death. NQO1 has been observed to be overexpressed in several types of solid tumors, including breast, pancreas, lung and colon cancer.
Shown to activate quinone based anti-tumor agents in vivo. Suitable for conjugation to carrier molecules.

Unit Definition

One unit will reduce 1.0 μmole cytochrome C per min/mg in the presence of menadione substrate at 37 °C.

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Eeva-Riikka Vehniäinen et al.
Aquatic toxicology (Amsterdam, Netherlands), 116-117, 102-108 (2012-04-10)
Ethoxyresorufin-O-deethylase (EROD) activity is a biomarker of exposure to planar aromatic hydrocarbons, and it is often measured from the S9 fraction. The effect of the liver S9 fraction of seven boreal freshwater fish species on the fluorescence of resorufin was
Molecular of Modelling of Human DT-Diaphorase for Enzyme-Directed Bioreductive Drug Design.
Doughtyl, S.W. and Phillips, R.M.
Molecular Simulations, 24, 209-209 (2000)
C Flader et al.
Journal of medicinal chemistry, 43(16), 3157-3167 (2000-08-24)
A series of naphthoquinone and benzimidazolequinone phosphorodiamidates has been synthesized and studied as potential cytotoxic prodrugs activated by DT-diaphorase. Reduction of the quinone moiety in the target compounds was expected to provide a pathway for expulsion of the phosphoramide mustard
Yang Yang et al.
Journal of experimental & clinical cancer research : CR, 33, 14-14 (2014-02-07)
NAD (P) H: quinone oxidoreductase 1 (NQO1) is a xenobiotic metabolizing enzyme that detoxifies chemical stressors and antioxidants, providing cytoprotection in normal tissues. However, high-level expression of NQO1 has been correlated with numerous human malignancies, suggesting a role in carcinogenesis
S Chen et al.
The Journal of biological chemistry, 272(3), 1437-1439 (1997-01-17)
DT-diaphorase (EC 1.6.99.2), also referred to as NAD(P)H:(quinone-acceptor) oxidoreductase, is involved in the reductive activation process of several cytotoxic antitumor quinones and nitrobenzenes. It has been observed in our and other laboratories that the rat enzyme is significantly more effective

Questions

1–6 of 6 Questions  
  1. Despite the recommended use of fresh solutions, can I store the NQO1 solution? What temperature and for low long?

    1 answer
    1. Unfortunately, the solution stability of this product has not been determined. In such situations, preparing solutions fresh is recommended to assure optimum performance. However, various sources suggest that solutions of DT Diaphorase containing 10% glycerol and/or 0.1% BSA may be stable for several months when stored in aliquots at -20°C. This information has not been validated.

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  2. Is it possible to obtain the concentration of α-Lactose monohydrate?

    1 answer
    1. The specific composition of the product is proprietary, so we cannot disclose the concentration of α-Lactose monohydrate. However, the DT Diaphorase protein accounts for approximately 5% of the total solids in the vial.

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  3. What is the molecular weight of this product?

    1 answer
    1. The product D1315 (DT Diaphorase (NQO1) human) is a monomer with a molecular weight of 30.87 KDa and has a homodimer structure.

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  4. What is the buffer composition of the lyophilized NQO1?

    1 answer
    1. The lyophilized product D1315 is prepared in PBS buffer with α-Lactose monohydrate sourced from bovine milk.

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  5. 본 제품인 DT-diaphorase (NQO1) human 의 수용해성 여부에 대한 정보가 없어 질문드립니다. 본 효소 제품이 수용해성을 가지고 있나요?

    1 answer
    1. This product is soluble in water at 1 mg/ml. It is recommended that solutions be freshly prepared before each use.

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  6. I already received this enzyme (DT-Diaphorase) for my experiments. I would like to put in a buffer it is suitable in. Would you have any information on this matter? Thanks!

    1 answer
    1. Product D1315 is lyophilized in PBS buffer with α-Lactose monohydrate. Since the powder already contains buffer and a stabilizer, it is recommended to use pure water. PBS is also an suitable option. It is recommended that the end-user does NOT try to weigh out a portion of the powder, because a representative sample may not be obtained. Please reconstitute the entire contents of the vial.

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