Annexin V is a calcium-dependent phospholipid-binding protein that specifically binds phosphatidylserine (PS) Normally PS is localized to the inner leaflet of the cell membrane lipid bilayer but during early stages of apoptosis PS flips from the inner leaflet to the outer surface of the plasma membrane where it is exposed to the extracellular environment Annexin V displays a high affinity for PS Fluorescein (FITC)-conjugated Annexin V can be used as a fluorescent probe in flow cytometry or fluorescent microscopy to detect apoptosis within 10-20 minutes The nuclear dye 7-Aminoactinomycin D (7-AAD) specifically binds to double-stranded DNA and fluoresces upon binding Under normal circumstances 7-AAD is cell-impermeable however in late apoptotic and dead cells 7-AAD can pass through cell membranes staining DNA Combining Annexin V-FITC with 7-AAD staining cells at different stages of apoptosis and dead cells can be distinguished effectively