【摘要】 为建立流式细胞术检测细胞内的信号转导及转录活化因子 3 (STAT3 )的方法 ,用IL 5刺激HL 60细胞株 ,活化细胞内的STAT3 ,然后用抗酪氨酸磷酸化STAT3抗体及相应的二抗标记 ,进行流式分析。结果表明 :( 1)IL 5能活化HL 60细胞内的STAT3 ,且STAT3的酪氨酸磷酸化水平与IL 5的浓度之间存在剂量依赖关系 ;( 2 )可用流式细胞术测得的平均荧光强度反映细胞内酪氨酸磷酸化STAT3的相对浓度。应用流式细胞术能快速、定量地测定细胞内的磷酸化STAT3水平。本法可用于STAT3相关疾病的筛选和临床监测及对其他胞内信号转导分子的研究。更多还原

【Abstract】 To establish a method of detecting the intracellular signal transduction and the phosphorylated STAT3 by flow cytometry,the HL 60 cell line was stimulated with IL 5 to activate the intracellular STAT3, and then measured with flow cytometry by anti tyrosine phosphorylated STAT3 antibody and the correpondding anti antibody labeling The results showed that this method could be used for rapid and quantitative assessment of STAT3 phosphorylation on a discrete cell basis and it proved to be more sensitive and less time consuming than immunoblotting Application of this method should be valuable in screening for signaling defects in selected patients with leukemia,cancers and so on In addition,this technique should permit dissection of a full range of cellular signaling pathways at the protein level更多还原