考马斯亮兰G—250染料测定蛋白质方法的改进
IMPROVEMENT OF PROTEIN DETERMINATION WITH COOMASSIE BRILLIANT BLUE G-250 DYE
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摘要: 考马斯亮兰G-250(简称G-250)酸性溶液呈红棕色,与蛋白质结合时变为兰色,A_(max)从465nm移至595nm。在一定条件下,蛋白质浓度与A_(595nm)成正比,据此作蛋白质定量。本法有快速、简便、灵敏和抗多种化学试剂干扰等优点,已得到广泛使用。但此法对不同的蛋白质定量有时差异较大,并时有不稳定的染料蛋白复合物沉出,影响测定准确度~4。作行用正交试验法选出了试剂配制的最佳条件,对经典法作了改进。Abstract: The effects of concentration of several reagents and time of color de velopment in coomassie brilliant blue G-250 assay on determination of five proteins were tested by orthogonal design. The optimum concentration of dye, ethanol and phosphoric acid were
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