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 37ºCand 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 asanticoagulan