Linearization of the bradford protein assay increases its sensitivity: Theoretical and experimental studies
Linearization of the bradford protein assay increases its sensitivity: Theoretical and experimental studies
复制标题
DOI：
10.1006/abio.1996.0171
复制
发表时间：
1996-05-01
影响因子：
2.9
通讯作者：
Seliger, Z
中科院分区：
文献类型：
--
作者：
Zor, T;Seliger, Z
通过测量590 nm处的吸光度，在Bradford考马斯亮蓝试验中测定微克量的蛋白质。然而，固有的非线性损害了该方法的灵敏度和准确性。结果表明，在标准测定条件下，590 nm与450 nm的荧光强度之比与蛋白质浓度呈严格线性关系。这个简单的程序增加了准确度，并提高了约10倍的测定灵敏度，允许定量低至50 ng的牛血清白蛋白。此外，可以在十二烷基硫酸钠（去污剂）相对于牛血清白蛋白（蛋白质）重量过量高达35倍的情况下进行蛋白质测定。根据质量作用和比耳定律，给出了与实验数据完全符合的线性方程。(C)出版社：Academic Press，Inc.
Determination of microgram quantities of protein in the Bradford Coomassie brilliant blue assay is accomplished by measurement of absorbance at 590 nm. However, an intrinsic nonlinearity compromises the sensitivity and accuracy of this method. It is shown that under standard assay conditions, the ratio of the absorbances, 590 nm over 450 nm, is strictly linear with protein concentration. This simple procedure increases the accuracy and improves the sensitivity of the assay about 10-fold, permitting quantitation down to 50 ng of bovine serum albumin. Furthermore, protein assay in presence of up to 35-fold weight excess of sodium dodecyl sulfate (detergent) over bovine serum albumin (protein) can be performed. A linear equation that perfectly fits the experimental data is provided on the basis of mass action and Beer's law. (C) 1996 Academic Press, Inc.