Principle
Manual assay procedure Prewarm at 37°C the required amount of working solution be fore use. Perform the assay as given below :
Creatine kinase catalyzes the conversion of creatine phosphate and ADP to creatine and ATP. ATP phosphorylates glucose to glucose-6-phosphate in the presence of hexokinase. Glucose-6-phosphate is oxidized to 6-phosphogluconate, reducing NADP to NADPH in presence of G-6-PDH. The rate of increase in NADPH absorbance at 340 nm. is directly proportional to the activity of creatine kinase in serum/plasma.
Procedure
Reaction type ………… UV – Kinetic
Reaction direction ………….. …………… Increasing
Wavelength ……… ………………… 340 nm.
Flow Cell temperature ………. ….37°C
Zero setting with …………………………………… Distilled water
Delay time ……….. ……. …………………….. 180 seconds
No. of readings ……………. …………………………4
Interval ……….. … …. …….. 30 seconds
Sample volume …………… ……………… …………0.04 ml (40 ul)
Working solution volume (4R, : 1R,) ……. 1.0 ml (1000 ul)
Factor ……………………….. 4127 0
Linearity. ………………… 2000 IU/
Quality Control
To ensure adequate quality control, it is recommended that each batch should include a normal and an abnormal commercial reference control serum. It should be realized that the use of quality control material checks both instrument and reagent functions together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glassware and accuracy of pipetting
Other Details
The reagent kit should be stored at 2 -8° C and is stable till the expiry date indicated on the label. R1 & R2 reagents are stable till expiry at 2 -8° C. The working solution (4 R1 + 1 R2) is stable for 10 days at 2 – 8° C.
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