Kit rises recombinant annexin V conjugated to allophycocyanin (APC), C12-resazurin and Sytox green stain and a simple method is used to stain the cell population
The new Vybrant apoptosis assay kit 10 from Molecular Probes provides a rapid and convenient assay for apoptosis.
It comprises recombinant annexin V conjugated to allophycocyanin (APC), C12-resazurin and Sytox green stain and a simple method is used to stain the cell population.
Apoptotic cells show far-red fluorescence, intermediate orange fluorescence and no green fluorescence.
Those in late apoptosis exhibit intense far-red and green fluorescence and little orange fluorescence.
Conversely, live cells show little or no green or far-red fluorescence, but there is significant fluorescence in the orange channel.
These populations can easily be distinguished using a flow cytometer. In apoptotic cells phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the plasma membrane exposing it to the external environment.
The human vascular anticoagulant, annexin V, is a 35-36 kDa Ca2+-dependent phospholipid-binding protein with a high affinity for PS.
Annexin V labelled with a fluorophore or biotin can identify apoptotic cells by binding to PS exposed on the outer leaflet.
Sytox green is impermeant to live and early apoptotic cells but stains late-stage apoptotic cells with intense green fluorescence.
Nonfluorescent C12-resazurin is reduced by viable cells to orange-fluorescent C12-resorufin.
Resazurin has been used extensively to detect the metabolic activity of many different cell types, from bacteria to higher eukaryotes.
