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Diaphorase (Escherichia coli)

Product code: E-DIAEC
€0.00

1,000 Units

Prices exclude VAT

This product has been discontinued

Content: 1,000 Units
Shipping Temperature: Ambient
Storage Temperature: 2-8oC
Formulation: In 3.2 M ammonium sulphate
Physical Form: Suspension
Stability: Minimum 1 year at 4oC. Check vial for details.
Enzyme Activity: Other Activities
EC Number: 1.8.1.4
CAS Number: 9001-18-7
Synonyms: dihydrolipoyl dehydrogenase; protein-N6 (dihydrolipoyl)lysine:NAD+ oxidoreductase
Source: Escherichia coli
Molecular Weight: 51,700
Concentration: Supplied at ~ 275 U/mL
Expression: Recombinant from Escherichia coli
Specificity: Catalyses the reaction:
Tetrazolium compound + NAD+ →  formazan compound + NADH + H+
Specific Activity: ~ 12 U/mg (25oC, pH 9.0)
Unit Definition: One Unit of diaphorase activity is defined as the amount of enzyme required to oxidise one µmole of NADH per minute with associated reduction of INT in Tris.HCl buffer (32.3 mM), pH 9.0 at 25oC.
Temperature Optima: 25oC
pH Optima: 9
Application examples: Used widely in diagnostic and analytical applications.

This product has been discontinued (read more).

High purity recombinant Diaphorase (Escherichia coli) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.

Looking for more enzyme products? See our complete list of analytical enzymes.

Documents
Certificate of Analysis
Safety Data Sheet
Data Sheet
Publications
Megazyme publication

A novel enzymatic method for the measurement of lactose in lactose‐free products.

Mangan, D., McCleary, B. V., Culleton, H., Cornaggia, C., Ivory, R., McKie, V. A., Delaney, E. & Kargelis, T. (2018). Journal of the Science of Food and Agriculture, 99, 947-956.

Background: In recent years there has been a surge in the number of commercially available lactose‐free variants of a wide variety of products. This presents an analytical challenge for the measurement of the residual lactose content in the presence of high levels of mono‐, di‐, and oligosaccharides. Results: In the current work, we describe the development of a novel enzymatic low‐lactose determination method termed LOLAC (low lactose), which is based on an optimized glucose removal pre‐treatment step followed by a sequential enzymatic assay that measures residual glucose and lactose in a single cuvette. Sensitivity was improved over existing enzymatic lactose assays through the extension of the typical glucose detection biochemical pathway to amplify the signal response. Selectivity for lactose in the presence of structurally similar oligosaccharides was provided by using a β-galactosidase with much improved selectivity over the analytical industry standards from Aspergillus oryzae and Escherichia coli (EcLacZ), coupled with a ‘creep’ calculation adjustment to account for any overestimation. The resulting enzymatic method was fully characterized in terms of its linear range (2.3-113 mg per 100 g), limit of detection (LOD) (0.13 mg per 100 g), limit of quantification (LOQ) (0.44 mg per 100 g) and reproducibility (≤ 3.2% coefficient of variation (CV)). A range of commercially available lactose‐free samples were analyzed with spiking experiments and excellent recoveries were obtained. Lactose quantitation in lactose‐free infant formula, a particularly challenging matrix, was carried out using the LOLAC method and the results compared favorably with those obtained from a United Kingdom Accreditation Service (UKAS) accredited laboratory employing quantitative high performance anion exchange chromatography - pulsed amperometric detection (HPAEC‐PAD) analysis. Conclusion: The LOLAC assay is the first reported enzymatic method that accurately quantitates lactose in lactose‐free samples.

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Safety Information
Symbol : Not Applicable
Signal Word : Not Applicable
Hazard Statements : Not Applicable
Precautionary Statements : Not Applicable
Safety Data Sheet
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