Wako Product Information
<For Research Use Only>
Wako Catalog No. 036-13393

Creatine Kinase (CK), from bovine heart



Assay:

  1. Principle:

  2. Phosphocreatine + ADP --(Creatine Kinase)--> Creatine + ATP
    ATP + Glucose --(HK)--> G6P + ADP
    G6P + NADP --(G6PDH)--> NADPH + 6PG
  3. Reagents

  4. A: 0.15mol/L Imidasol buffer (pH7.4)
    B: Substrate solution:
    <Preparation>
    Dissolve the following reagents into Reagent A and adjust each concentration as mentioned below: C: Hexokinase (1,000 units/mL)
    D: Glucose 6-Phosphate Dehydrogenase (1,000 units/mL)
    E: Enzyme solution
    12 mg of this product is dissolved to Reagent A to prepare the approximately 3 units/mL solution.
  5. Procedure
  6. Reagent
    Test
    Control
    B
    3.00 mL
    3.00 mL
    Preincubate at 30°C
    C
    0.01 mL
    0.01 mL
    D
    0.01 mL
    0.01 mL
    E
    0.02 mL
    -
    A
    -
    0.02 mL
    Record the absorbance change at 340 nm after 2 ~ 5 minute incubation at 30°C and calculate the average change in absorbance per minute (‡™A) according to JIS K0115.
  7. Unit Definition

  8. One unit is defined as the amount of enzyme to produce 1 ƒÊmol of NADPH per minute at 30°C and pH 7.4.
  9. Calculation

  10. unit/vial = [(‡™A test - ‡™A control ) X 3.04] / ( 6.2 X S X 0.02 )


Product:

Description
Package Size
Wako Catalog No.
Storage
Creatine Kinase (CK), from bovine heart
2,000 units
036-13393
Keep at -20°C

Reference:
Dawson, D. M. and Eppenberger, H. M.: Meth. Enzymol., 17A, 995 (1970).


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