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作 者:范世光[1] 顾鹏毅[1] 赵蔓嘉 汪佳慧[1] 张潇木[1] 刘国栋[1] 梁再赋[1] 孙晋民[1]
机构地区:[1]中国医科大学实验技术中心一部暨国家中医药管理局中药生物工程实验室Ⅲ级,沈阳110001
出 处:《西北药学杂志》2015年第4期334-337,共4页Northwest Pharmaceutical Journal
基 金:辽宁省科技厅课题项目(编号:2013225021);辽宁省科技厅课题项目(编号:2008225021-1)
摘 要:目的探讨微孔板酶联比色测定法在蚯蚓粗提物蛋白质量浓度、蚓激酶活性测定中的应用。方法蛋白质量浓度采用改良Lowry法,蚓激酶活性采用TAME法,分别使用常量体积和微量体积,用紫外-可见分光光度计和酶标检测仪进行测量,用SPSS软件进行直线拟合,做出标准曲线,比较测量结果。结果标准曲线截距RSD<5%,用标准曲线检测BSA的结果显示,2种质量浓度、2种方法的回收率分别为99.6%,94.6%,101.2%和96.5%,RSD分别为3.30%,5.05%,3.36%和2.59%,经t检验3种检测结果差异无统计学意义,可以认为2种检测方法测得的数据一致。结论可以采用微量法进行蚯蚓粗提物蛋白质量浓度、蚓激酶活性的测定。Objective To investigate and compare the application of microplate enzyme-linked colorimetric assay in protein concentration determination,and the kinase activity in earthworm crude extracts.Methods The protein concentration was determined by using the improved Lowry method,and the kinase activity was determined by using the TAME method.The constant volume and micro volume were tested respectively,in UV-VIS spectrophotometer or enzyme measurement standard detector.SPSS software was used to make the standard curve.Results The standard curve intercept RSD was less than 5%.Two concentration curves was used for BSA detection.The recovery of the two methods were 99.6%,94.6%,101.2% and 96.5%,respectivly,and RSD were3.30%,5.05%,3.36% and 2.59%,respectivly.No statistical significance was observed.Conclusion The microplate assay method could be used for the protein concentration and the kinase activity determination.
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