Shishido K, Sudo J
Department of Toxicology and Clinical Pharmacology, Faculty of Pharmaceutical Sciences, Higashi-Nippon-Gakuen University, Hokkaido, Japan.
J Toxicol Sci. 1988 Feb;13(1):1-6. doi: 10.2131/jts.13.1.
In this study, the original method (Lowry method) of Lowry et al. (1951), its modified method (Lowry-TCA method) with protein precipitation by trichloroacetic acid (TCA) and the new method (BCA method) with bicinchoninic acid reaction, were reexamined as to whether these three methods were applicable to urinary protein quantitation of rats. Because of the presence of the urinary interfering substances, the quantitations in the Lowry- and BCA methods were found to be overestimated more than 3 times as large as in the Lowry-TCA method. When the urine sample was dialyzed against water and determined by the Lowry- and BCA methods, more than 24 hours were found to be needed in order to completely exclude the interfering substances from the urine. From these findings, the Lowry-TCA method is recommended for quantitation of urinary protein in rats.
在本研究中,对Lowry等人(1951年)的原始方法(Lowry法)、用三氯乙酸(TCA)沉淀蛋白质的改良方法(Lowry-TCA法)以及采用二辛可宁酸反应的新方法(BCA法)进行了重新检验,以确定这三种方法是否适用于大鼠尿蛋白定量。由于存在尿干扰物质,发现Lowry法和BCA法的定量结果比Lowry-TCA法高估了3倍以上。当尿样用水透析并用Lowry法和BCA法测定时,发现需要超过24小时才能完全排除尿中的干扰物质。基于这些发现,推荐使用Lowry-TCA法对大鼠尿蛋白进行定量。