Flow cytometry has traditionally been used to detect extracellular proteins to identify different cell populations. With the advancement of technology and reagents in recent years, we now are able to easily detect intracellular proteins in individual cells. One very useful application for the detection of intracellular proteins is to measure specific cells signaling pathways. These pathways are often altered in disease states, such as cancer, and are now easily detected using flow cytometry providing single cell analysis of these signaling changes.
Jurkat cells were either left in culture(pERK unstim) or stimulated with cell stimulation cocktail for 1hr (pERK +stim). Cells were washed, fixed and permeablized, followed by staining with anti-phospo-ERK1/2 (T204/Y202) PerCP antibody for 1hr. Cells were washed and analyzed on the NovoCyte flow cytometer for phospho-ERK levels and compared using a histogram overlay generated with the NovoExpress acquisition and analysis software.