
UNIT DEFINITION
One unit (U) is defined as the amount of enzyme which consumes 1 μmol of NADH per min under the conditions described below.
CAS no. | 9028-36-8 |
REAGENTS PREPARATION
Reagent I: Add 0.04 g sodium pyruvate in 0.1 M, pH 7.7 Tris-HCl buffer and set to 100 mL.
Reagent II: Add 2.11 g CAPS in 80 mL pure water and adjust to pH 9.7, then add 0.12 g NADH and set to 100 mL.
Enzyme diluent: 10 mM, pH 7.0 potassium phosphate, contains 0.1% BSA.
Sample: The enzyme was diluted to 1-2 U/mL with Enzyme diluent.
PROCEDURE
- Add 1mL Reagent I and 0.02mL enzyme solution to the cuvette.
- Preincubate the reaction mixture at 37 ℃ for 5 min.
- Add 0.2 mL Reagent II and mix, record the ΔAs at 340 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 in stead of the enzyme solution.
∆A = ∆As – ∆Ab

df: dilution factor
C: Enzyme concentration (mg/mL)
6.22: Millimolar extinction coefficient of NADH under 340 nm (cm2/μmol)
REFERENCES
- C.A. Loshon, R.B. McComb, L.W. Bond, G.N. Bowers, Jr.W.H. Coleman and R.H.Gwynn; Clin. Chem., 23, 1576 (1977).
- H. Taguchi, M. Machida, H. Matsuzawa and
- Ohta; Agric. Biol. Chem., 49 (2), 359 (1985).
- F. Gasser, M. Doudoroff, and R. Contopoulos;
- Gen. Microbiol. 62, 241 (1970).
This enzyme is used for the removal of pyruvate in determinations working with NADH (triglycerides, lipase, aldolase, aspartate aminotransferases, glutamate dehydrogenase)

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