Detection of Both Surface Proteins and Intracellular Phospho-Determinants in Whole Blood Using the PerFix-p Reagent System
Flow cytometry provides a unique methodology to monitor signaling pathways, allowing a coordinated measurement of multiple phosphoepitopes in the context of cell surface markers. Application of this technology has been hindered by the diversity of sample preparation protocols designed to unmask particular epitopes, often at the expense of surface marker detection. A novel sample preparation methodology, the PerFix-p* reagent system (Beckman Coulter), allows simultaneous and reproducible detection of multiple phospho-epitopes in conjunction with surface markers on a variety of samples, including whole blood, peripheral blood mononuclear cells, and bone marrow. The ability to multiplex phospho-epitope detection with immunophenotyping opens new possibilities to map changes in the phosphorylation pathways associated with cellular processes in normal and abnormal cells.