Reference: DAD-301

Diaphorase

Diaphorase from Clostridium NAD(P)H + H+ + Acceptor (ox) ----> NAD(P)+ + Acceptor (red) This enzyme is used for colorimetric determination of NAD(P)H and many dehydrogenases when coupled with various dyes which act as hydrogen acceptors from NAD(P)H.
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DESCRIPTION
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  • Name: NAD(P)H (acceptor) oxydoreductase
  • Appearance: Yellowish amorphous powder lyophilized
  • Activity: Grade III 30U/mg-solid or more (containing approx. 15% of stabilizers)
  • Contaminants: Myokinase ≤ 5.0×10-1%
    NAD(P)H oxidase ≤ 5.0×10-1%
  • Stabilizers: FMN, NAD(P)H
  • Stability: Stable at -20 °C for at least one year
  • Molecular weight: 24,000
  • Michaelis constants: 2.0×10-5 M (NADH), 6.0×10-6 M (NADPH)
  • Structure: One mol of FMN per mol of enzyme
  • Inhibitor: N-Ethylmaleimide
  • Optimum pH: 8.5
  • Optimum temperature: 50 °C
  • pH Stability: pH 7.5 (30 °C, 3hr)
  • Thermal stability: below 30 °C (pH 7.5, 30min)
  • Substrate specificity: Either NADH or NADPH can be used as a reductant. The ratio (NADPH/NADH) is 0.6 in the assay method. Neither oxygen nor cytochrome C can be utilized as a hydrogen acceptor.
  • EC # 1.6.99.-

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