微量法在蚯蚓粗提物蛋白质量浓度和蚓激酶活性测定中的应用

摘要

Objective To investigate and compare the application of microplate enzyme‐linked colorimetric assay in protein concentra‐tion determination ,and the kinase activity in earthworm crude extracts .Methods The protein concentration was determined by u‐sing 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 were 3 .30% ,5 .05% ,3 .36% and 2 .59% ,respectivly .No statistical significance was observed .Conclusion The microplate assay meth‐od could be used for the protein concentration and the kinase activity determination .%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。