二喹啉甲酸检测法测定Sprague-Dawley大鼠肝微粒体蛋白浓度

Study on Bincinchonicic Acid Methodology of Detecting Protein Concentration in Liver Microsomes of Sprague-Dawley Rats

目的建立酶标仪检测大鼠肝微粒体蛋白浓度的方法。方法大鼠尾静脉连续3 d注射0.9%氯化钠注射液2 ml,第4 d解剖大鼠,取肝脏,应用差速离心法提取大鼠肝微粒体,用二喹啉甲酸检测（BCA）法测定肝微粒体蛋白浓度。结果蛋白标准曲线范围在0~0.5mg/ml,回归方程为Y=0.091 9＋0.868X,（R=0.998）。低、中、高浓度回收率分别为101.33%,110.67%,101.17%,日内和日间精密度分别为6.52%,0.53%,1.95%和3.01%,7.23%,0.74%。结论本实验建立的BCA蛋白测定法准确可靠,重复性和稳定性均良好,相比较传统的蛋白测定方法,更为迅速,具有较强的抗干扰的能力,为研究用药后肝微粒体酶活性的改变提供可靠的检测手段。

Objective To establish the bincinchonicic acid（BCA） protein assay methodology in liver microsomes of Sprague-Dawley（SD） rats.Methods After treated with 0.9% NaCl once a day for 3 d,the SD rats were anatomized to get liver microsomes with different velocity centrifugations.The BCA method was used to measure the concentration of protein in liver microsomes.Results There was a good linearity within the range of 0~0.5 mg/ml between the concentration and absorbance.The regression equation was Y=0.091 9＋0.868X（R=0.998）.The recoveries were 101.33%,110.67%,101.17%（n=3） respectively,intra-day precision were 6.52%,0.53%,1.95% respectively and inter-day precision were 3.01%,7.23%,0.74% respectively.Conclusion The BCA protein assay method was accurate,sensitive,stable,rapid and repeatble compared with traditional detecting methods,and it provided a credible way for protein assay.