[Objective]This study was to develop a detection method for assaying calf intestine alkaline phosphatase activity in scientific research. [Method]By simulating the conditions and buffers for assaying calf intestine alkaline phosphatase used in the scientific research,the parameters influencing substrate concentration,reaction duration,coloration time and buffer p H were optimized. [Result]The activity of alkaline phosphatase detected varied hugely among different buffers,and potassium ferricyanide solution added with boracic acid achieved the stable coloration. The optimal water bath time was determined as 10 min,and the substrate concentration was optimized as 0. 04 mol / L. The increasing temperature did not have a large influence on low temperature enzymatic activity while did on high concentration enzymatic activity. When the buffer p H was 7. 0- 8. 0,the detection stability of alkaline phosphatase could be maintained well. The ion intensity of buffer had little effects on alkaline phosphatase activity. [Conclusion]A detection method for assaying calf intestine alkaline phosphatase activity in scientific research was successfully developed in this study.
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