陕西医学杂志2011，Vol.40Issue(3)：259-261,279,4.
非同位素标记DNA探针和凝胶迁移分析法测定NF-κB激活实验研究
Detection of NF-κB with non-radioactive labeled DNA probes and gel shift assays
摘要
Abstract
Objective: To study the advantages of detection of nuclear factor-κB(NF-κB) with digoxingenin-labeled DNA probes and gel shift assays. Methods : DNA probes were labeled with digoxingenin and labeling efficiency was determined. Following the binding reaction of DNA probes and nuclear protein, the complexes were resolved with polyacrylamide gel electrophoresis (PAGE), and transferred to a positively charged nylon membrane. The membrane was dried, incubated with alkaline phosphatase-conjugated anti-digoxingenin antibody and then detected with CSPD. Results : The labeling efficiency was high. Nuclear protein was combined with digoxigenin-labeled DNA probes and showed a high density shift band after gel shift assays.Competition experiments using unlabeled NF-κB or control Oct2A DNA probes and supershift assays using specific antibodies against P65 and P50 demonstrated the specificity of the DNA protein binding reaction.Conclusion: This method has the advantages of better stability, sensitivity and safety without radioactive contamination and is easy to operate. The pictures are clear with low background.关键词
NF-κB/分析/凝胶迁移分析实验/DNA探针/利用分类
医药卫生引用本文复制引用
牛红心..非同位素标记DNA探针和凝胶迁移分析法测定NF-κB激活实验研究[J].陕西医学杂志,2011,40(3):259-261,279,4.基金项目
广东省自然科学基金项目(编号9451051501002540)广东省科技计划项目(编号2010B031600239) （编号9451051501002540）
