一种核蛋白提取方法在血管内皮细胞NF-κB活性研究中的作用

Role of an approach for nucleic protein extracts from vascular endothelial cells in the detection of nuclear factor-κB

目的探讨改良核蛋白提取方法在血管内皮细胞核因子-κB(NF-κB)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-κB活性。结果提取的血管内皮细胞核蛋白样品蛋白含量约2.5 g·L-1,受刺激核蛋白样品中有明显的NF-κB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-κB活性好,适合大量细胞核蛋白提取和进一步对样品中NF-κB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-κB（NF-κB）. Methods Extracting solutions were prepared and nucleic protein of vascular endothelial cells were extracted.Sample protein concentration was detected with Bradford method and NF-κB activity was detected with gel shift assay and Western blotting. Results Sample protein concentration reached up to about 2.5 g·L-1.NF-κB 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-κB activity in the nucleic extracts,thus is suitable for the nucleic protein extraction from a large number of cells and further research for NF-κB activity.