Biosystems Creatinkinaz Reagent
PRINCIPLE OF THE METHOD
Creatine kinase (CK) catalyzes the phosphorylation of ADP, in the presence of creatine
phosphate, to form ATP and creatine. The catalytic concentration is determined from the rate of
NADPH formation, measured at 340 nm, by means of the hexokinase (HK) and glucose-6-
phosphate dehydrogenase (G6P-DH) coupled reactions1,2
A. Reagent: Imidazol 125 mmol/L, EDTA 2 mmol/L, magnesium acetate 12.5 mmol/L, Dglucose 25 mmol/L, N-acetyl cysteine 25 mmol/L, hexokinase 6000 U/L, NADP 2.4 mmol/L,
B. Reagent: Creatine phosphate 250 mmol/L, ADP 15 mmol/L, AMP 25 mmol/L, P1,P5-
di(adenosine-5′-)pentaphosphate, 102 µmol/L, glucose-6-phosphate dehydrogenase
Store at 2-8ºC.
Reagents are stable until the expiry date shown on the label when stored tightly closed and if
contaminations are prevented during their use.
Indications of deterioration:
− Reagents: Presence of particulate material, turbidity, absorbance of the blank over 0.300 at
340 nm (1 cm cuvette).
Working Reagent: Pour the contents of the Reagent B into the Reagent A bottle. Mix gently.
Other volumes can be prepared in the proportion: 4 mL Reagent A + 1 mL Reagent B.
Stable for 15 days at 2-8ºC. The working reagent must be protected from light.
− Analyzer, spectrophotometer or photometer with cell holder thermostatable at 25, 30 or 37ºC
and able to read at 340 nm.
− Cuvettes with 1 cm light path.
Serum and plasma collected by standard procedures.
Creatine kinase in serum and plasma is stable for 7 days at 2-8ºC. Use heparin or EDTA as