 | Principle of the Assay
The Muse™ Annexin V and Dead Cell Assay is based on the detection of phosphatidylserine (PS) on the surface of apoptotic cells, using fluorescently labeled Annexin V in combination with the dead cell marker, 7-AAD. Annexin V is a Ca2+-dependent phospholipid binding protein that has a high affinity for PS, a membrane component normally localized to the internal face of the cell membrane. Early in the apoptotic pathway, molecules of PS are translocated to the outer surface of the cell membrane where Annexin V can readily bind to them. Late-stage apoptotic cells show loss of membrane integrity. The membrane-impermeant dye 7-AAD is used to distinguish dead cells from early apoptotic cells. The assay can thus distinguish four populations:
- Viable cells, not undergoing detectable apoptosis:
Annexin V (–) and dead cell marker (–)
- Early apoptotic cells: Annexin V (+) and dead
cell marker (–)
- Late apoptotic cells: Annexin V (+) and dead cell
marker (+)
- Cells that have died through non-apoptotic
pathway: Annexin V (–) and dead cell marker (+)
Muse™ Annexin V and Dead Cell Assay Features
- Quick determination of live cells, early and late
apoptotic cells, and dead cells
- No-wash, mix-and-read assay
- Highly simplified acquisition and analysis
- Minimal number of cells required
- Validated with both adherent and suspension cells
- Accurate and precise
Click the PDF above for the full document. |
