The RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay measures the real-time exposure of phosphatidylserine (PS) on the outer leaflet of cell membranes during the apoptotic process. Annexin V luciferase fusion proteins supplied in the assay reagent bind to PS during early apoptosis and are detected with a simple luminescence signal. The assay reagent also includes a DNA-binding dye, which enters the cell and generates a fluorescent signal upon loss of membrane integrity.
The combination and timing of luminescent (annexin V binding) and fluorescent (DNA release) signals is used to differentiate secondary necrosis occurring during late apoptosis from necrosis caused by other cytotoxic events.
The RealTime-Glo™ Annexin V Apoptosis and Necrosis assay is non-lytic and the simple “add-and-read” method allows multiple readings from a single assay well. Apoptosis can be monitored in real time, without the need for multiple plates, complicated processing, or specialized detection equipment. A multimode reader capable of detecting luminescence and fluorescence is the only instrument required.