【求助】有谁用流式细胞术检测过蛋白磷酸化吗?

有谁用流式细胞术检测过蛋白磷酸化吗？
我现在做信号通路研究，要用PE标记的单抗检测细胞内的信号分子磷酸化状态，涉及到透膜处理，具体的标记的实验步骤以及用什么buffer有谁知道吗？

蛋白磷酸化为什么不用western检测？

Are you using the BD cell signalling master kit?
If so, I think you need to prepare the cell lysate instead of the permeablization.

P. Nolan in the Department of Microbiology and Immunology, School of Medicine, Stanford UniversityGarry is the No.1 expert on this aspect. You can logon his group to download some of their paper and protocol.

Attached please find the protocol of intracellular phopho-protein detecting by FACS