【求助】用流式细胞仪检测JAK-STAT5通路？
丁香园论坛
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有谁用流式细胞仪检测过STAT-5磷酸化程度呀，求教具体方法，结果怎么样呀？
1、制备样品细胞的单细胞悬液。
2、多聚甲醛固定细胞。
3、打孔剂在细胞膜上打孔。
4、标记荧光素偶联的抗磷酸化STAT-5抗体。
5、流式上样分析。
Crucial role of interleukin-7 in T helper type 17 survival and expansion in autoimmune disease. Nature Medicine. 2010. 16(2):191-197
这篇文章有用到该法。文章可以到《免疫学与病原生物学讨论版》置顶贴中找。
2、多聚甲醛固定细胞。
3、打孔剂在细胞膜上打孔。
4、标记荧光素偶联的抗磷酸化STAT-5抗体。
5、流式上样分析。
Crucial role of interleukin-7 in T helper type 17 survival and expansion in autoimmune disease. Nature Medicine. 2010. 16(2):191-197
这篇文章有用到该法。文章可以到《免疫学与病原生物学讨论版》置顶贴中找。
kern6549 wrote:
1、制备样品细胞的单细胞悬液。
2、多聚甲醛固定细胞。
3、打孔剂在细胞膜上打孔。
4、标记荧光素偶联的抗磷酸化STAT-5抗体。
5、流式上样分析。
Crucial role of interleukin-7 in T helper type 17 survival and expansion in autoimmune disease. Nature Medicine. 2010. 16(2):191-197
这篇文章有用到该法。文章可以到《免疫学与病原生物学讨论版》置顶贴中找。
这篇文章中的方法写得非常简单，能否介绍1篇步骤稍稍详细的文章或把以上步骤描述得详细一点，谢谢！
我上传几篇流式细胞仪检测蛋白磷酸化的详细说明，供大家实验参考
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