Hiratsuka N, Shiba K, Shinomura K, Hosaki S
School of Allied Health Sciences, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
J Clin Lab Anal. 1996;10(6):403-6. doi: 10.1002/(SICI)1098-2825(1996)10:6<403::AID-JCLA15>3.0.CO;2-0.
In order to establish a rapid staining method sensitive enough to stain the protein bands on cellulose acetate membrane and suitable for the analysis of urinary proteins in healthy subjects, we conducted a detailed examination of Kovarik's method designed for the analysis of proteins in cerebrospinal fluid in detail. For purpose, of our study, we changed the pH of the staining solution used in Kovarik's method to pH 4.4 by adding acetic acid. With this solution, our staining method revealed a degree of sensitivity about three times higher than that of the original method, but specific sensitivity for albumin and gamma-globulin remained almost unchanged. The solution preparation we used was much more simple and the reagent much more stable than in the original method. Our method also showed better reproducibility and linearity than the original method. These results point to the suitability of this method for the analysis of low-concentration proteins, such as urinary proteins, in healthy subjects.
为了建立一种灵敏度足够高、能对醋酸纤维素膜上的蛋白条带进行染色且适用于健康受试者尿蛋白分析的快速染色方法,我们对为脑脊液中蛋白质分析而设计的科瓦里克方法进行了详细研究。为此,我们通过添加乙酸将科瓦里克方法中使用的染色溶液的pH值调至4.4。使用该溶液,我们的染色方法显示出的灵敏度比原方法高约三倍,但对白蛋白和γ-球蛋白的特异性灵敏度几乎保持不变。我们使用的溶液制备比原方法简单得多,试剂也更稳定。我们的方法还比原方法显示出更好的重现性和线性。这些结果表明该方法适用于健康受试者中低浓度蛋白质(如尿蛋白)的分析。