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通过低温预冷处理有机溶剂,分级提取兔肝碱性磷酸酶,结合SDS-PAGE法检测纯化结果。对酶的比活力测定中,蛋白定量经过对Bradford法和Folin-Lowry法进行比较,发现Folin-Lowry法在本样品体系中灵敏度高、抗干扰能力强,适合采用此法进行蛋白含量。经过对酶提取条件、酶的分子鉴定、比活力测定中蛋白含量检测方法的优化,使实验的实践性、应用性得到拓展。
The organic solvent was precooled at low temperature, the rabbit hepatic alkaline phosphatase was fractionated and purified by SDS-PAGE. Compared with Bradford method and Folin-Lowry method, it was found that Folin-Lowry method has high sensitivity and strong anti-interference ability in this sample system, which is suitable for the determination of protein content. After the enzyme extraction conditions, the molecular identification of enzymes, specific activity determination of protein content in the detection method optimization, the experiment of practicality, applicability has been expanded.