固体表面荧光技术在蛋白质定量分析中的应用
首发时间:2009-09-24
摘要:以硅胶为固相基质材料和邻苯二甲醛为荧光探针,初步研究了固体表面荧光技术在蛋白质定量分析中的应用。用单因素法对实验参数进行了优化,确定最佳试剂浓度、试剂固化时间和检测酸度分别为0.6mg mL-1、1.0min和pH 12.0。在最佳实验条件下,可对1~200μg mL-1浓度范围内的蛋白质进行准确定量分析,检出限为0.56μg mL-1,分析精度为1.6%(50μg mL-1, n=11)。对人血清总蛋白进行分析测定,测定结果与考马斯亮蓝方法一致;牛奶样品中总蛋白测定结果加标加收率为98.9%.
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Investigations on the determination of proteins with solid-phase surface fluorescence
Abstract:A solid-phase surface fluorescence technique was developed for the determination of protein by exploiting silica gel as solid phase and o-phthaldialdehyde(OPA) as fluorescence probe. A uni-variant approach was adopted for the optimization of the experimental parameters, resulting in the use of a 0.6mg mL-1 OPA, an immobilization time of 1 min and a sample acidity of pH 12 for the quantitative analysis. A linear calibration graph was obtained in a range of 1冾200
Keywords: solid-phase surface fluorescence silica gel protein o-phthaldialdehyde
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