Diaphorase CAS UENA-0247

Identification

CAS Number

UENA-0247

Name

Diaphorase

EC

1.6.5.2

Organism species

Microorganism

Source

Recombinant expressed by E.coli

Formulation

≥160U/mg yellowish amorphous lyophilized power. Refer to the COA and the bottle logo, or according to customer’s special requirement.

Molecular mass

~26kDa by SDS-PAGE

Properties

Appearance

Pale yellow amorphous lyophilized powder

Safety Data

RIDADR 

NONH for all modes of transport

Specifications and Other Information of Our Diaphorase CAS UENA-0247

Protein purity

>90% by SDS-PAGE (Fig 1).

Unit definition

One unit is defined as the amount of enzyme which oxidizes one micromole NADH per minute under the conditions of pH7.0 at 37℃ as described.The activity concentration (U/L) is calculated by detecting ∆A/min of formazane dye production rate at 550nm optical wavelength. Millimolar extinction coefficient of formazane dye under the assay conditions is 12.4 cm2/micromole.

Enzyme properties

1) Inhibitor: Sodium azide can be used as a preservative for enzyme solution and lyophilized powder without affecting activity and stability, but 0.1% sodium azide in the detection Reagent R1 and Reagent R2 can inhibit the activity about 25%.
2) Stabilizer and cofactor: FMN.
3) Optimum pH: pH7.0 (Fig 3).
4) pH stability: pH5.0~10.0 (Fig 4).
5) Optimum temperature: above 60℃ (Fig 5).
6) Thermal stability: below 70℃ in PBS, pH7.4 (Fig 6).
7) Freeze-Thaw Stability: after 20 freeze-thaw cycles, the activity of liquid was stable (Fig 7).
8) Accelerated stability: The activity of enzyme solution and lyophilized power are stable by incubated at 42℃ for 3 days and 37℃ for 14 days (Fig 8 and 9).
9) Special activity: ~400U/mg protein.
10) Km=0.22mM (NADH).

Storage

Stable at -20℃ for at least 12 months (Fig 2).

Known Application

As an indicator enzyme, DPR is useful for enzymatic determination of D-3-Hydroxybutyrate when coupled with D-3-Hydroxybutyrate Dehydrogenase (HBDH, EC 1.1.1.30) in clinical analysis. DPR is also useful for enzymatic determination of NADH and other dehydrogenases.

This product is developed by our R&D company Watson International Ltd (https://www.watson-int.com/).