考马斯亮蓝(CBB)显色法测定蛋白质含量的主要缺点之一是线性关系差.通过研究显色液组分对线性关系的影响,发现显色液H+浓度是影响线性关系的主要因素,并提出了一个新的显色液配方来改善考马斯亮蓝蛋白质测定法的线性关系.
The coomassie brilliant blue (CBB) protein assay first developed by Bradford became an alternative for an ever-increasing number of researchers because of its simplicity, rapidity and being free from interference by some common laboratory reagents which have been shown to affect the lowry assay. Despite all its advantages, many problems have been encountered by researchers in using this procedure. One of the main drawbacks of this procedure is the nonlinearity which has been noted since Bradford developed the CBB-protein assay. The effect of ingredients in color-developing reagent on the absorbances at 465 nm and 595 nm, of free CBB and protein-binding CBB mixture was tested respectively. The results showed that the main factor which affects the linearity of calibration curve is H+ concentration. A new color-developing reagent formula which consists of CBB, HCl and NaCl, may improve the nonlinearity.
郭敏亮,姜涌明.考马斯亮蓝显色液组分对蛋白质测定的影响[J].生物化学与生物物理进展,1996,23(6):558-561
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