BD Phosflow Cell Signaling

Researchers can use BD Phosflow™ technology to distinguish and analyze phosphoprotein signaling in single cells through the use of multiple cell surface markers.


BD Phosflow™ allows scientists to narrow down to small cellular subsets in complex samples, such as whole blood or peripheral blood mononuclear cells (PBMCs). Unlike traditional methods such as Western blotting, researchers can use BD Phosflow to distinguish and analyze phosphoprotein signaling in single cells through the use of multiple cell surface markers. Even rare cell subtypes can be uncovered and isolated without additional upfront methods.

The ability to study complex events under near-native conditions provides greater accuracy for understanding the effect of a disease or stimulus. For drug discovery, BD Phosflow is also a unique secondary, cell-based screening tool for cell types such as mouse splenic cells.

By enabling investigators to analyze phosphorylation state in whole blood or other complex cell mixtures, BD Phosflow helps streamline discovery. It eliminates the need for lengthy and costly experiments designed to reconcile unexpected effects stemming from rare cell populations or discrepancies between in vivo and in vitro results.


Human whole blood analysis with BD Phosflow.

Human whole blood analysis with BD Phosflow

In this experiment, human whole blood was activated using anti-CD3, anti-CD28, and anti-CD49d in the presence of increasing concentrations of the tyrosine kinase inhibitor imatinib mesylate (Gleevec®). Phospho-ERK expression was measured in CD4+, and CD4- T cells after 5 minutes of stimulation and compared with IL-2 and IFN-γ intracellular cytokine expression after a 5-hour incubation with the activating antibody cocktail. The percentage of cells that expressed phospho-ERK or IL-2 was decreased in both CD4+ and CD4- T cell populations in the presence of increasing concentrations of Gleevec. However, the number of IFN-γ expressing cells was less affected by Gleevec in CD4- T cells.

Human whole blood analysis with BD Phosflow.

In this experiment, human whole blood was activated using anti-CD3, anti-CD28, and anti-CD49d in the presence of increasing concentrations of the tyrosine kinase inhibitor imatinib mesylate (Gleevec®). Phospho-ERK expression was measured in CD4+, and CD4- T cells after 5 minutes of stimulation and compared with IL-2 and IFN-γ intracellular cytokine expression after a 5-hour incubation with the activating antibody cocktail. The percentage of cells that expressed phospho-ERK or IL-2 was decreased in both CD4+ and CD4- T cell populations in the presence of increasing concentrations of Gleevec. However, the number of IFN-γ expressing cells was less affected by Gleevec in CD4- T cells.