Intracellular Flow Cytometry Tips And Tricks

Flow cytometry protocols and staining procedures vary depending on whether the antigen to be detected is located on the cell surface or intracellular. Detection of cell surface proteins, such as CD markers, is relatively straightforward and apart from blocking of FcR receptors requires no extra protocol steps. However, when staining intracellular proteins, such as cytokines or transcription factors, both fixation and permeabilization steps are required prior to antibody staining.

Read on to learn tips such as carefully choosing the most suitable fixative and permeabilization reagents as well as other factors that should be considered when designing intracellular flow experiments.