摘要
免疫荧光可定性分析细胞内蛋白核转位现象,NF-κB/p65蛋白核转位提示NF-κB信号通路的活化,我们尝试运用自主研发设计的免疫荧光分析软件计算荧光图像中胞核和胞质位置的荧光值变化,定量分析核转位情况,并通过蛋白质免疫印迹法验证软件分析结果。应用该软件分析内毒素诱导0.5、1、2和4h后原代人脐静脉内皮细胞的NF-κB信号通路活化,结果提示2h为核转位高峰期,与蛋白质免疫印迹法验证结果一致,表明自主研发的免疫荧光分析软件可应用于免疫荧光的定量分析。
Immuno-fluorescence technique can qualitatively determine certain nuclear translocation,of which NF-κB/p65 implicates the activation of NF-κB signal pathways.Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm.We verified the quantitative analysis by Western Blot.When we applied the software to analysis of nuclear translocation in lipopolysaccharide(LPS)induced(0.5h,1h,2h,4h)primary human umbilical vein endothelial cells(HUVECs),we found that nuclear translocation peak showed up at 2has with calculated Western blot verification results,indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2015年第3期669-674,共6页
Journal of Biomedical Engineering
基金
国家自然科学基金资助项目(81160193)
江西省卫生厅科技计划资助项目(20121053)
关键词
免疫荧光
荧光定量分析
NF-ΚB
人脐静脉内皮细胞
内毒素
immuno-fluorescence
fluorescence quantitative analysis
NF-kappa B
human umbilical vein endothelialcells
lipopolysaeeharide