温度对考马斯亮蓝法测定蛋白质浓度的影响

Effects of Temperature on Determination of Protein Concentration with Coomassie Brilliant Blue Method

[目的]研究温度在考马斯亮蓝法测定蛋白质过程中的影响,为准确测定蛋白质浓度提供建议和指导。[方法]通过考马斯亮蓝法测定在不同温度和孵育时间下对已知不同浓度的牛血清蛋白的影响。[结果]在0-100 mg/ml蛋白质浓度标准曲线测定范围内,室温20℃孵育20-30 min时测定的蛋白质浓度比较稳定。同时蛋白质浓度越高,考马斯亮蓝法测定蛋白质浓度过程中对各种因素影响的抵御能力越强。[结论]采用考马斯亮蓝法测定蛋白质浓度时,环境温度、待测样品浓度以及染料与蛋白质孵育时间等是决定实验结果准确性的重要因子。

[Objective] This study aimed to investigate the effects of temperature on determination of protein concentration with coomassie brilliant blue method,thus proving advice and guidance for accurate determination of protein concentration.[Method] With coomassie brilliant blue method,the concentrations of different bovine serum albumin samples were determined under different temperature and incubation time.[Result]When the determination range of protein concentration was 0-100 mg / ml according to the standard curve,the determined protein concentration was relatively stable after incubation at 20 ℃ for 20-30 min. Furthermore,the determination result of higher protein concentration with coomassie brilliant blue method was less affected by various factors. [Conclusion] In determination of protein concentration with coomassie brilliant blue method,the temperature,sample concentration and incubation time were important factors affecting the accuracy of experimental results.