详细信息:Millipore’s FlowCellect™ MitoStress Kit includes MitoSOX Red, a live-cell permeant fluorogenic dye which targets the mitochondria and reacts with superoxide radicals and fluoresces yellow/red, and Annexin V conjugated to CF647 which binds to phosphatidylserine (PS) on the surface of apoptotic cells. The simultaneous use of these reagents allows researchers to obtain information on oxidative stress and apoptosis in one simple assay.
MitoSOX Red is excitable by a 488nm laser and fluoresces maximally at 580 nm (Yellow fluorescence on the guava easyCyte 8HT). Annexin V is calcium-dependent phospholipid binding protein with high affinity for PS, a membrane component normally localized to the internal face of the cell membrane. It is conjugated to CF647 which is excited by a red laser and emits at 665 nm (Red2 fluorescence on the guava easyCyte 8HT). Control cells depict no Red2 fluorescence while apoptotic cells exhibit positive Red2 fluorescence. The kit can thus distinguish multiple populations: 1) Live cells with little or no superoxide 2) Stressed cells with oxidized MitoSOX Red (indicator of accumulated superoxide) but no Annexin V staining 3) Stressed and Early Apoptotic cells with oxidized MitoSOX Red and Annexin V binding and 4) Apoptotic Cells with Annexin V binding only., The MitoStress kit rapidly provides information on correlation of mitochondrial superoxide stress with apoptosis using minimal cellular sample. The entire assay can be performed in less than 30 minutes, once cellular samples are ready.
