
Unit definition
One unit is defined as the amount of enzyme which produces 1 μmol of H2O2 per minute under the conditions described below.
CAS 9028-72-2
Reagents preparation
Reagent I:0.2 M pH 6.5 potassium phosphate buffer.
Reagent II: 1kU/mL peroxidase (POD) solution.
Reagent III:50 mM 4-AA solution.
Reagent IV: 0.5 M DL- Lactic acid solution, pH 6.5.
Reagent V:50 mM TOOS solution.
Enzyme diluent : 10 mM pH 7.0 potassium phosphate solution with 10 μM FAD. to 0.05 – 0.2 U/mL.
Prepare reaction mixture as follows:
Reagent I 10 ml
Reagent II 0.25 mL
Reagent III 1.5 mL
Reagent IV 5 mL
Reagent V 1.5 mL
Add ddH2O to 50 ml
Procedure
- Add 1 mL of the reaction mixture to 1 mL cuvette.
- Heat the reaction mixture at 37 °C for 5 min.
- Add 20 μL of the enzyme solution to the cuvette and mix.
- Record the ΔAs at 555 nm in 1 minute in a spectrophotometer thermo stated 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

1.02: Total volume of reaction solution (mL)
0.02: Volume of enzyme solution (mL)
1: Light path length (cm)
1/2: 1mol H2O2 will react to 1/2 mol
Quinoneimine dye df: Dilution multiple
C: Enzyme concentration (mg/mL)
39.2: Millimolar extinction coefficient of quinoneimine dye under 555nm (cm2/μmol)
This enzyme is useful for enzymatic determination of lactic acid.

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