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Fructosyl-Peptide Oxidase
- Supports IFCC traceable enzymatic assays;
- Features superior specificity and accuracy, without interference from subtypes of hemoglobin A (HbA);
- Maintains superior stability, stable for 18 months if tightly sealed and stored at <4°C and for 28 days after opening;
- Displays high intra- and inter-assay precision, with the intra-assay CV ≤4% and the inter-assay CV ≤6%.
Fructosyl-peptide oxidase (EC 1.5.3, from Microorganism)
Fructosyl-peptide + H2O + O2 ⇒ Peptide + Glucosone + H2O
he assay is based on the increase in absorbance at 555 nm as the formation of quinoneimine dye proceeds in the forward reactions.
UNIT DEFINITION
One unit (U) is defined as the amount of enzyme which produces 1 μmol of H2O2 per min under the conditions described below.
REAGENTS PREPARATION
Reagent I: 0.1 M potassium phosphate, pH 8.0.
Reagent II: 1 kU/mL POD.
Reagent III: 50 mM TOOS solution.
Reagent IV: 50 mM 4-AA solution.
Reagent V: 200 mM fructosyl-valine solution.
Enzyme diluent: 20 mM Tris-HCl,pH 8.0.
Sample: Dilute the sample to 0.2-0.5 U/mL by Enzyme diluent.
Reaction mixture:
Reagent I 10 mL
Reagent II 0.1 mL
Reagent III 1 mL
Reagent IV 1 mL
Reagent V 10 mL
Pure water Set to 100mL
PROCEDURE
- Add 0.98 mL reaction mixture to the cuvette.
- Preincubate the reaction mixture at 37 ℃ for 5 min.
- Add 0.02 ml of the enzyme solution to the reaction mixture.
- 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
Vt: Total volume (1.0 mL)
Vs: Enzyme volume (0.02 mL)
t: reaction time (1 min)
df: dilution factor
C: Enzyme concentration (mg/mL)
1.0: Light path length (cm)
1/2: 1mol H2O2 will react to 1/2 mol
Quinoneimine dye
39.2: Millimolar extinction coefficient of
quinone imine dye under 555nm (cm2/μmol)
The enzyme is useful for the determination of fructosyl-peptide and fructosyl-L-amino acid.
The Certificate Of Analysis (COA) & Material Safety Data Sheet (MSDS) Is A Signed Document That Includes The Storage Temperature,
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