
UNIT DEFINITION
One unit (U) is defined as the amount of enzyme which consumes 1 μmol of DCPIP per min under the conditions described below.
REAGENTS PREPARATION
Reagent I: 200mM pH7.0 Tris-HCl buffer.
Reagent II: 36 mM NADH solution.
Reagent III: 2.4 mM DCPIP solution.
Enzyme diluent: 200 mM Tris-HCl, pH7.5, contains 0.5% Tween20.
Sample: dilute the enzyme to 0.1-0.25 U/ml with enzyme diluent.
PROCEDURE
- Add 2.4ml pure water, 0.3ml Reagent I and 0.1ml Reagent II into a 3ml cuvette and equilibrate at 37 ℃ for about 4 minutes.
- Add 0.1 mL of samples and 0.1 mL DCPIP to mix.
- Record the ΔAs at 600 nm in 1 minute in a spectrophotometer thermostated at 37℃.
At the same time, measure the blank rate ΔAb by using the same method as the test except that the enzyme diluent is added instead of the enzyme solution.
∆A = ∆As – ∆Ab

20.9: Millimolar extinction coefficient of DCPIP under the assay conditions (cm2/μmol)
REFERENCES
1) F. Kaplan, P. Setlow and N.O. Kaplan; Arch, Biochem. Biophys., 132, 91 (1969).
This enzyme is useful for colorimetric determination of NAD(P)H and many dehydrogenases when coupled with various dyes which act as hydrogen acceptors from NAD(P)H.

The Certificate Of Analysis (COA) & Material Safety Data Sheet (MSDS) Is A Signed Document That Includes The Storage Temperature,
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