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目的建立淋巴细胞内磷酸化信号传导及转录激活子(STAT)的快速检测方法。方法人单个核细胞或外周静脉血淋巴细胞用IL-6、IL-4和IFN-α1体外刺激15min,运用流式细胞术检测胞内STAT3(pY705)、STAT6(pY641)和STAT1(pY701)磷酸化水平,并比较不同破膜剂对它的影响。结果与0.2%皂素、90%甲醇比较,PurmbufferⅢ破膜剂破膜后,能够显著地提高淋巴细胞内STAT3的磷酸化水平(P<0.01)。结论经PermbufferⅢ破膜后,运用流式细胞术可快速检测磷酸化STAT3、STAT6和STAT1。
Objective To establish a rapid detection method of phosphorylation signal transducers and activators of transcription (STAT) in lymphocytes. Methods Human mononuclear cells or peripheral blood lymphocytes were stimulated with IL-6, IL-4 and IFN-α1 in vitro for 15 min. Flow cytometry was used to detect intracellular STAT3 (pY705), STAT6 (pY641) and STAT1 (pY701) Level, and compare the effect of different rupture agents on it. Results Compared with 0.2% saponin and 90% methanol, Purmbuffer Ⅲ rupture membrane could significantly increase the phosphorylation of STAT3 in lymphocytes (P <0.01). Conclusion Permbuffer Ⅲ membrane rupture, the use of flow cytometry can be quickly detected phosphorylation of STAT3, STAT6 and STAT1.