MEASUREMENT OF PROTEIN USING BICINCHONINIC ACID
MEASUREMENT OF PROTEIN USING BICINCHONINIC ACID
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DOI：
10.1016/0003-2697(85)90442-7
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发表时间：
1985-01-01
影响因子：
2.9
通讯作者：
KLENK, DC
中科院分区：
文献类型：
--
作者：
SMITH, PK;KROHN, RI;KLENK, DC
联苯胆酸，钠盐，是一种稳定的、水溶性的化合物，能够在碱性环境中与Cu1+形成强烈的紫色络合物。该试剂构成了一种分析方法的基础，该方法能够监测蛋白质与碱性Cu2+反应(双缩二脲反应)产生的Cu2+。这种反应产生的颜色是稳定的，并且在蛋白质浓度增加的大范围内按比例增加。与Lowry等人的方法相比，本文报告的结果表明，该试剂对非离子洗涤剂和简单缓冲盐等常见干扰有更大的耐受性。试剂和所产生的发色团的稳定性还允许简化的一步分析和增强方案选择的灵活性。这种新方法保持了与Lowry技术相关的高灵敏度和低蛋白质对蛋白质的变化。
Bicinchoninic acid, sodium salt, is a stable, water-soluble compound capable of forming an intense purple complex with cuprous ion (Cu1+) in an alkaline environment. This reagent forms the basis of an analytical method capable of monitoring cuprous ion produced in the reaction of protein with alkaline Cu2+ (biuret reaction). The color produced from this reaction is stable and increases in a proportional fashion over a broad range of increasing protein concentrations. When compared to the method of Lowry et al., the results reported here demonstrate a greater tolerance of the bicinchoninate reagent toward such commonly encountered interferences as nonionic detergents and simple buffer salts. The stability of the reagent and resulting chromophore also allows for a simplified, one-step analysis and an enhanced flexibility in protocol selection. This new method maintains the high sensitivity and low protein-to-protein variation associated with the Lowry technique.