NF-κB是一种诱导性转录因子, 广泛存在于各种细胞, 在细胞受到各种刺激时可被激活, 调节大量靶基因, 因此在很多重要细胞进程, 如细胞生长、分化、凋亡和癌变中均发挥重要作用。自其发现以来的25年中, NF-κB是生物医学科学各领域中被持续大量深入研究的前沿热点转录因子, 同时也是疾病治疗和药物筛选的重要靶点。NF-κB的活性检测是研究其活化和功能首当其冲的实验内容, 因此NF-κB的活性检测技术历来受到重视和不断发展。特别是近几年来, 随着各学科的发展, 出现了多种NF-κB活性检测新技术, 如基于双链DNA修饰微孔板的类ELISA分析、膜结合分析、各种荧光共振能量转移分析、基于内外切酶保护的荧光报告及核酸扩增分析、基于免疫微球的质谱及流式细胞分析, 以及物理化学分析等。其中有些技术已经对NF-κB的研究发挥了重要作用。文章对近年来发展的各种检测技术进行了分类综述, 以便从事NF-κB相关研究的科研人员对该种转录因子的检测分析技术有一个全貌的了解, 并有益于在其研究中选择恰当合适的实验方法。此外, 对这些技术的学习和理解, 可能激发研究人员对现有技术的改进和发展新的技术。
NF-κB is a stimulatory transcription factor that is ubiquitous in almost all kinds of cells. When cells are under various stimuli, NF-κB is activated and regulates large numbers of target genes, and thus controls important cellular processes, ranging from cell growth and differentiation to apoptosis and cancer. Therefore, NF-κB is a forefront hotspot transcription factor that is intensively studied in virtually all fields of biomedical sciences, and becomes a promising target for disease therapy and drug development. The activity detection is the first and inevitable step for the studies of NF-κB activation and function, therefore, the techniques for the detection of NF-κB activity have always been paid more attention and continuously developed. Especially in recent year, along with the development of each disciplines, multiple various new techniques have been developed, including ELISA-like assays based on dsDNA-coupled plate, filter binding assays, FRET assays, fluorescence reporting and nucleic acids amplification assays based on exonuclease and endonuclease, MS and flow cytometry assays based on immunomicrobeads, and other biophysical and electrochemical assays. Some of these techniques have already played important roles in NF-κB studies. This paper reviewed new techniques developed in recent years by classification, in order to provide researchers an overview of NF-κB activity assays, which may be helpful for their selection of appropriate techniques for their studies. Moreover, the learning and understanding of these techniques may inspire researchers to improve the current techniques and develop novel methods for the studies of NF-κB.
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