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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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一种简便的高通量细胞内gfp报告基因检测方法
Keywords: 绿色荧光蛋白,报告基因,高通量检测,多功能酶标仪
Abstract:
？目的:建立一种简单、快速、高通量的检测gfp蛋白的方法。方法:用tecan多功能酶标仪infinitem200pro检测gfp蛋白在不同缓冲液中的荧光强度;用酶标仪检测含gfp的蛋白样品梯度稀释后的荧光强度,建立荧光强度与gfp相对含量之间的标准曲线;分别比较荧光酶标仪的检测结果与流式细胞术和western-blotting检测结果的相关性。结果:使用50mmol/ltris-hcl(ph=10)的缓冲液稀释gfp蛋白更有利于检测低浓度的gfp;荧光酶标仪检测gfp的荧光读数同gfp相对含量之间的回归系数r2>0.99。荧光酶标仪检测gfp蛋白的结果与流式细胞术、western-blotting的检测结果之间均呈现明显的相关性。结论:建立的利用多功能酶标仪检测gfp蛋白方法方便快捷,可以实现短时间对大样本进行检测。
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