This kit contains CF488A Annexin V for staining apoptotic cells green, and 7-AAD for staining necrotic cells with far-red fluorescence, for detection by flow cytometry or fluorescence microscopy.
Phosphatidylserine (PS) is translocated from the inner to the outer surface of apoptotic cells for phagocytic cell recognition. The human anticoagulant, Annexin V, is a 35 kDa Ca2 -dependent phospholipid protein with a high affinity for PS. Annexin V labeled with CF488A (Abs/Em= 490/515 nm) can identify apoptotic cells in green by binding to PS exposed on the outer leaflet. CF488A dye is superior to fluorescein/FITC as it is not affected by pH and has much better photostability.
7-AAD is a membrane-impermeant nucleic acid dye that is excluded by live cells, but stains necrotic cells and late apoptotic cells. 7-AAD (Abs/Em with DNA = 543/655 nm) emits in the far-red spectrum and can be well separated from CF 488A and other green emitting dyes using the 488 nm line of a flow cytometer.
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