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通过一种普遍适用的比浊法测定微克量的蛋白质。

Measurement of microgram quantities of protein by a generally applicable turbidimetric procedure.

作者信息

Vera J C

机构信息

Instituto de Bioquímica, Universidad Austral de Chile, Valdivia.

出版信息

Anal Biochem. 1988 Oct;174(1):187-96. doi: 10.1016/0003-2697(88)90534-9.

Abstract

A modified turbidimetric method for protein determination which involves the use of trichloroacetic acid as the precipitating agent is described. Maximal turbidity develops in less than 30 min and is stable for at least 120 min. A linear relationship between turbidity at 340 nm and protein concentration is observed between 2 and 40 micrograms protein. Sodium dodecyl sulfate is added to avoid the interference by nonionic and cationic detergents and lipids and to decrease the protein-to-protein variation. The use of cetyltrimethyl ammonium bromide provides a two-step procedure to correct for the contribution of contaminating nucleic acid. Many compounds which interfere with other protein quantitation methods have no effect on this system. The interference of commonly used reagents as sucrose and urea can be easily corrected. This procedure compared favorably with the most widely used protein quantitation methods in simplicity, sensitivity, and specificity.

摘要

本文描述了一种改良的比浊法用于蛋白质测定,该方法使用三氯乙酸作为沉淀剂。最大浊度在30分钟内形成,并至少稳定120分钟。在2至40微克蛋白质之间,观察到340nm处的浊度与蛋白质浓度呈线性关系。添加十二烷基硫酸钠以避免非离子和阳离子洗涤剂及脂质的干扰,并减少蛋白质间的差异。使用十六烷基三甲基溴化铵提供了一个两步程序来校正污染核酸的贡献。许多干扰其他蛋白质定量方法的化合物对该系统没有影响。常用试剂如蔗糖和尿素的干扰可以很容易地校正。该方法在简便性、灵敏度和特异性方面与最广泛使用的蛋白质定量方法相比具有优势。

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