目的 探讨改良核蛋白提取方法在血管内皮细胞核因子-kB(NF-kB)活性研究中的作用及其优势.方法 配制核蛋白提取液,提取血管内皮细胞核蛋白,用Bradford法测定样品蛋白含量,用凝胶迁移实验和Western blotting 检测NF-kB活性.结果 提取的血管内皮细胞核蛋白样品蛋白含量约2.5g·L-1,受刺激核蛋白样品中有明显的NF-kB激活.结论 本方法操作简便、快捷、经济,所提取的核蛋白,蛋白含量高,NF-kB活性好,适合大量细胞核蛋白提取和进一步对样品中NF-kB活性的分析研究.%Objective To explore the role and advantages of an improved approach for nucleic protein extracts from vascular endothelial cells in the detection of nuclear factor-KB ( NF-kB ). Methods Extracting solutions were prepared and nucleic protein of vascular endothelial cells were extracted. Sample protein concentration was detected with Bradford method and NF-kB activity was detected with gel shift assay and Western blotting. Results Sample protein concentration reached up to about 2.5 g · L-1. NF-kB showed obviously activated in the stimulated nucleic protein extracts. Conclusion This method has the advantages of simpler operation, lower prices and higher protein concentration and NF-kB activity in the nucleic extracts, thus is suitable for the nucleic protein extraction from a large number of cells and further research for NF-kB activity.
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