1.   Samples should not contain Fe3+ and Cu2+, or it will inhibit the activity of G-6-PD.

2.   Avoid moderate or severe hemolysis of samples. When the degree of hemolysis is high, red blood cells will release AK, ATP, G-6-PD and so on, which will affect the results.

3.   The activity of CK is unstable, temperature and light may lead to the loss of enzyme activity. So the sample should be detect as soon as possible after collection, or preserve the samples on ice for before detection.

4.   When the reagent 1 is taken, it is recommended to avoid the contamination of reagent.

1)   Blank tube: Take 1000 μL of reagent 1, 50 μL of double distilled water into an EP tube.

Sample tube: Take 1000 μL of reagent 1, 50 μL of sample into EP tubes.

2)   Mix fully and incubate at 37℃ for 5 min.

3)   Add 100 μL of preheated reagent 2.

4)   Mix fully and incubate at 37℃ for 2 min.

5)   Set to zero with blank tube and measure the absorbance at 340 nm with 1 cm quartz cuvette at initial absorbance (A1) and 5 min (A2), respectively. Calculate the △A=A1-A2.