三种酶纯化常用试剂对Bradford法的干扰研究  

Study on the interference of three common reagents for enzyme purification on Bradford's assay

作　　者：孙欣欣 张兴国 伍校军 汤欢 宋晓梅 陈同 邓祥宜 SUN Xinxin ZHANG Xingguo WU Xiaojun TANG Huan SONG Xiaomei CHEN Tong DENG Xiangyi(College of Food and Biological Science and Technology, Wuhan Institute of Design and Sciences, Wuhan 430205, China College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China)

机构地区：[1]武汉设计工程学院食品与生物科技学院,武汉430205 [2]华中农业大学食品科技学院,武汉430070

出　　处：《黑龙江畜牧兽医》2017年第10期278-281,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金面上项目(31471428);湖北省大学生科技创新项目(201414035007)

摘　　要：为了研究常用酶纯化试剂对考马斯亮蓝法(Bradford法)测定蛋白质含量的影响,试验以牛血清白蛋白(BSA)为标准蛋白,探索硫酸铵、乙醇和丙酮对该方法的干扰规律和消除这些干扰的方法,然后在丙酮沉淀脂肪酶试验中进行验证。结果表明:硫酸铵对蛋白质含量测定值影响较小,而高浓度的丙酮、乙醇造成测定值偏高数倍;将蛋白质样品稀释5倍可基本消除丙酮、乙醇的干扰。在验证试验中,将上清液稀释5倍,然后测定蛋白质含量,解决了随着丙酮浓度升高,沉淀和上清液蛋白质含量测定值同时增加的问题。说明稀释是解决高浓度乙醇、丙酮对Bradford法干扰的有效方法。In order to investigate the effect of common reagents for enzyme purification on Bradford＇ s assay in determining protein content, bo- vine serum albumin（BSA） served as a standard protein in exploring interference rules of ammonium sulfate, ethanol and acetone on Bradford＇s assay, and exploring the scheme of interference elimination, and verification of the scheme through acetone - used lipase precipitation trial. The results showed that ammonium sulfate took a little effect on protein content determination,but high concentrations of acetone or ethanol caused the determining values several times larger than the actual values. Five - fold dilution of protein sample could eliminate nearly the interference of acetone or ethanol. In verification trial, five - fold dilution of the supernatant in protein content determination, solved the issue that the deter- mining values of protein content in both precipitate or supernatant increased simultaneously with the increase of acetone concentration. The re- sults indicated that dilution is an effective method to eliminate the interference of high concentrations of ethanol or acetone on Bradford＇ s assay.

关 键 词：考马斯亮蓝法(Bradford法) 蛋白质含量 丙酮 乙醇 干扰 测定 

分 类 号：S854.43[农业科学—临床兽医学]